KR102503593B1 - Composition for diagnosing or treating drug resistant cancer - Google Patents

Abstract

The present invention relates to a composition for treating anticancer drug resistance that diagnoses and overcomes anticancer drug resistance using a novel biomarker composition for diagnosing anticancer drug resistance. It is expected that the patient's cost burden can be reduced by predicting the suitability of each patient's anticancer drug before administration and providing accurate basic information, and at the same time, the survival rate can be improved by treating anticancer drug resistance.

Description

Composition for diagnosing or treating anticancer drug resistance {Composition for diagnosing or treating drug resistant cancer}

The present invention relates to a composition capable of diagnosing anticancer drug resistance and further treating the resistance.

Cancer is one of the incurable diseases that mankind must solve, and a huge amount of capital is being invested in development to cure it worldwide. Diagnosed, more than 60,000 people die. In particular, gastric cancer was the fifth most frequently diagnosed disease worldwide in 2018, and although various anticancer therapies have been rapidly developed in cancer diagnosis and treatment over the past decade, the fatality rate due to cancer is still high. In addition, there are still side effects associated with trying various anticancer drugs and various anticancer therapies. Studies to reduce these side effects are being actively conducted.

Drug resistance (drug resistance) is a major factor that halves the effect of successful anti-cancer treatment, and deteriorates the prognosis of various carcinomas. Drug resistance (chemo-resistance) in cancer cells is divided into pre-existing of resistance-mediating factor and newly acquired drug resistance (acquired drug resistance) due to drug administration (Cancer drug resistance). : an evolving paradigm. Nat Rev Cancer. 2013 Oct;13(10):714-26.). Factors that cause acquired drug resistance include, for example, increased drug efflux, mutagenesis of drug targets, DNA damage repair, activation of alternative signaling pathways, or evasion of cell death induced by drug resistance. As such, even if a drug-resistant patient is treated with the drug, the effect of the treatment cannot be guaranteed, and thus, there is a possibility that the time and cost of clinicians and patients are unnecessarily consumed. Therefore, when starting anticancer treatment, it is necessary to make a decision in consideration of the individual characteristics of the patient according to the treatment. A scale that exists is a realistic need. As such, since there is almost no research related to drug resistance, the present inventors have completed the present invention by discovering a novel marker capable of pre-selecting patients with anticancer drug resistance.

One object of the present invention is to provide a composition for diagnosing anticancer drug resistance.

Another object of the present invention is to provide a kit for diagnosing anticancer drug resistance.

Another object of the present invention is to provide a method for providing information on anticancer drug resistance.

Another object of the present invention is to provide a pharmaceutical composition for treating anticancer drug resistance.

Another object of the present invention is to provide a pharmaceutical composition that enhances sensitivity to anticancer drugs.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating anticancer drug-resistant cancer.

Another object of the present invention is to provide a screening method for a drug for overcoming or treating anticancer drug resistance or a drug for enhancing sensitivity to an anticancer drug.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein are described with reference to the drawings. In the following description, numerous specific details are set forth, such as specific forms, compositions and processes, etc., in order to provide a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well known processes and manufacturing techniques have not been described in specific detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, the appearances of "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, particular features, forms, compositions, or properties may be combined in one or more embodiments in any suitable way.

Unless there is a specific definition within the specification, all scientific and technical terms used herein have the same meaning as commonly understood by a person skilled in the art to which the present invention belongs.

1. Use of anticancer drug resistance diagnosis

According to one embodiment of the present invention, it relates to a composition for diagnosing anticancer drug resistance.

In the present invention, the diagnostic composition may include an agent for measuring the expression level of AcAT (acetylated alpha tubulin) protein or a gene encoding the same.

In the present invention, the term "acetylated alpha tubulin (AcAT)" is known to encode a mitochondrial localization enzyme that catalyzes the reversible formation of acetoacetyl-CoA from two molecules of acetyl-CoA by this gene. Causes of defects in the gene include 3-ketothiolase deficiency, congenital isoleucine catabolism characterized by urinary excretion of 2-methyl-3-hydroxybutyric acid, 2-methyl acetoacetic acid, tigliglycine and butanone. related to the error.

In the present invention, information about the "AcAT protein or gene encoding the same" is registered in NCBI (National Center for Biotechnology Information), the AcAT protein may consist of the amino acid sequence represented by SEQ ID NO: 1, and the AcAT protein The encoding gene may consist of the nucleotide sequence represented by SEQ ID NO: 2, but is not limited thereto, and as a non-limiting example, 99% or more to less than 100%, 95% or more to less than 99%, 90% or more of the AcAT sequence % or more to less than 95%, 85% or more to less than 90%, or more than 80% to less than 85% homology, and to those skilled in the art, it is known that the desired effect of the present invention is exerted. All may be included without limitation within the obvious range.

In the present invention, "medicine" or "anti-cancer treatment" and "anti-cancer agent" may be used interchangeably, and the drug corresponds to a drug having an anti-cancer effect by killing not only cancer stem cells but also cancer cells, and more preferably for gastric cancer treatment. refers to drugs that are effective.

In the present invention, the "anti-cancer agent" is a drug having a mechanism of killing cancer cells, including nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, Semasanib, bosutinib, axitinib, cediranib, restaurtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, bevacizumab, cisplatin, cetuximab, viscum album , asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine, gemtuzumabozogamicin, ibritumomabtusetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, pro Carbazine, alprostadil, holmium nitrate chitosan, gemcitabine, doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacitidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludabine, enocitabine, flutamide, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan , vinorelbine, etoposide, vincristine, vinblastine, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pirarubi Cin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, thiotepa, nimustine, chlorambucil , mitolactol, leucovorin, tretonin, exmestane, aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole , bicalutamide, lomustine, and may include one or more selected from the group consisting of carmustine, but is not limited thereto.

In the present invention, the "5-Fluorouracil" is a drug mainly used for breast cancer and digestive malignancies, and was developed as a structural analogue of thymine, a DNA precursor, by Heidelberger and Ansfield in 1957 . It belongs to an antimetabolic agent and corresponds to a drug having an action of inhibiting a metabolic process necessary for the proliferation of cancer cells. Drugs with similar actions include Cytarabine, Mercaptopurine (6-MP), and Methotrexate (MTX).

In the present invention, the "oxaliplatin" is an alkylating agent that modifies DNA structure and causes chain scission to correspond to a drug that exhibits anticancer and cytotoxic effects. Examples of drugs with similar actions include Busulfan, Chlorambucil, Cyclophosphamide, Melphalan, Nitrogen Mustard, Nitrosourea, These include Cisplatin and Ifosfamide.

In the present invention, the term "anti-cancer drug resistance" refers to a decrease in the effectiveness of a drug when the drug is used repeatedly in quantitative terms, and in order to obtain the same effect previously experienced by patients with anti-cancer drug resistance, the amount of use or the frequency of use must be increased. It is a condition in which the same effect cannot be obtained even if the substance needs to be increased or administered in the same amount as before. In the present invention, drug resistance was diagnosed when the change in the expression level of the AcAT protein or the gene encoding it was increased compared to the control group.

In the present invention, the "diagnosis" refers to determining susceptibility to an anticancer drug, determining whether an onset disease currently has anticancer drug resistance, or prognosis of an anticancer drug resistant cancer (e.g., cancer against that drug). determination of reactivity), or a broad concept that includes providing information on the diagnosis.

The "prognosis" of the present invention refers to the act of predicting the course of a disease and the outcome of death or survival in advance. The prognosis or diagnosis of prognosis may be interpreted as meaning that the course of a disease may vary depending on the patient's physiological or environmental state, and that it refers to all activities that predict the course of a disease before/after treatment by comprehensively considering the patient's condition. can For the purpose of the present invention, the prognosis is the act of predicting in advance whether treatment responsiveness or resistance to the anticancer drug will occur after treatment with an anticancer drug, preferably 5-fluorouracil or oxaliplatin, or whether or not the anticancer drug is used based thereon. It can be interpreted as an act of choosing appropriately.

In the present invention, "tumor" or "cancer" is a disease in which cell division continues due to unregulated cell cycle, and is divided into carcinoma and sarcoma according to the site of occurrence. Carcinoma refers to a malignant tumor arising from epithelial cells such as mucous membranes and skin, and sarcoma refers to a malignant tumor arising from non-epithelial cells such as muscle, connective tissue, bone, cartilage, and blood vessels. The cancers include thyroid cancer, parathyroid cancer, gastric cancer, ovarian cancer, colon cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, and hematological cancer. , bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, perianal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine adenocarcinoma, adrenal cancer , soft tissue sarcoma, urethral cancer, penile cancer, ureteric cancer, renal cell carcinoma, renal pelvic carcinoma, CNS central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma, but If the progression of cancer, such as differentiation and/or proliferation, is dependent on the cancer cells and/or cancer stem cells described in the present invention, it is not limited thereto.

In the present invention, "CD133" is an antigen also known as prominin-1 and corresponds to a glycoprotein encoded by the PROM1 gene in humans. It is a member of the pentaspan transmembrane glycoproteins of cell protrusions. As a marker specific to cancer stem cells, more specifically, the CD133 antigen refers to a cell-surface glycoprotein involved in cell regeneration, tumorigenesis, metastasis, metabolism, differentiation, and cell death. The CD133 biomarker can be used to identify cells with characteristics of cancer stem cells.

In the present invention, "CD44" is a marker expressed in the plasma membrane of cancer stem cells such as cells or drug-resistant cancer cells, and more specifically, the CD44 antigen is a cell-surface sugar involved in cell-cell interaction, cell adhesion and migration. say protein. CD44 is known to play a key role in tumorigenesis and plasticity of cancer stem cells.

The agent for measuring the expression level of the protein in the composition for diagnosing anticancer drug resistance of the present invention is from the group consisting of an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer that specifically bind to the protein It may include one or more selected species, but is not limited thereto.

In the present invention, the "antibody" refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, antibody means an antibody that specifically binds to said protein. Antibodies of the present invention include both polyclonal antibodies, monoclonal antibodies and recombinant antibodies. Such antibodies can be readily prepared using techniques well known in the art. For example, polyclonal antibodies can be produced by a method well known in the art, including a process of injecting an antigen of the protein into an animal and collecting blood from the animal to obtain serum containing the antibody. Such polyclonal antibodies can be prepared from any animal, such as goat, rabbit, sheep, monkey, horse, pig, cow, dog, and the like. In addition, monoclonal antibodies can be prepared using the hybridoma method well known in the art (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), or the phage antibody library technique (Clackson et al, Nature, 352). :624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). Antibodies prepared by the above methods may be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. In addition, the antibodies of the present invention include functional fragments of antibody molecules as well as complete forms having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule means a fragment having at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2, and Fv.

In the present invention, the "oligopeptide" is a peptide composed of 2 to 20 amino acids and may include a dipeptide, tripeptide, tetrapeptide and pentapeptide, but is not limited thereto.

In the present invention, the "PNA (Peptide Nucleic Acid)" refers to an artificially synthesized polymer similar to DNA or RNA, and was first introduced in 1991 by Professors Nielsen, Egholm, Berg and Buchardt of the University of Copenhagen, Denmark. Whereas DNA has a phosphate-ribose backbone, PNA has a repeated N-(2-aminoethyl)-glycine backbone linked by peptide bonds, which greatly increases the binding force and stability to DNA or RNA, and is thus used in molecular biology. , diagnostic assays and antisense therapies. PNA is described by Nielsen PE, Egholm M, Berg RH, Buchardt O (December 1991). "Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide". Science 254 (5037): 1497-1500.

In the present invention, the "aptamer" is an oligonucleic acid or peptide molecule, and a general description of the aptamer is described in the literature [Bock LC et al., Nature 355(6360):5646(1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630 (2000); Cohen BA, Colas P, Brent R. "An artificial cell-cycle inhibitor isolated from a combinatorial library". Proc Natl Acad Sci USA. 95(24): 142727 (1998).

In the composition for diagnosing anticancer drug resistance of the present invention, the agent for measuring the expression level of the gene encoding the protein comprises at least one selected from the group consisting of a primer, a probe, and an antisense nucleotide that specifically binds to the gene encoding the protein It may include, but is not limited thereto.

In the present invention, the "primer" is a fragment that recognizes a target gene sequence, and includes a forward and reverse primer pair, preferably a primer pair that provides an analysis result having specificity and sensitivity. High specificity can be imparted when the nucleic acid sequence of the primer is a sequence that is inconsistent with the non-target sequence present in the sample, so that the primer amplifies only the target gene sequence containing a complementary primer binding site and does not cause non-specific amplification. .

In the present invention, the "probe" means a substance that can specifically bind to a target substance to be detected in a sample, and means a substance that can specifically confirm the presence of a target substance in a sample through the binding. The type of probe is not limited as a material commonly used in the art, but preferably may be peptide nucleic acid (PNA), locked nucleic acid (LNA), peptide, polypeptide, protein, RNA or DNA, and most preferably Most likely it is PNA. More specifically, the probe is a biomaterial, including one derived from or similar to a living organism or manufactured in vitro, for example, enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, nerve cells, DNA, and It may be RNA, DNA includes cDNA, genomic DNA, and oligonucleotides, RNA includes genomic RNA, mRNA, and oligonucleotides, and examples of proteins may include antibodies, antigens, enzymes, peptides, and the like.

In the present invention, the "LNA (Locked nucleic acids)" means a nucleic acid analog containing a 2'-O, 4'-C methylene bridge [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502 ]. LNA nucleosides contain the common nucleic acid bases of DNA and RNA, and can base pair according to the Watson-Crick base pairing rules. However, due to molecular 'locking' due to the methylene bridge, the LNA does not form an ideal shape in the Watson-Crick bond. When LNAs are included in DNA or RNA oligonucleotides, they can more rapidly pair with complementary nucleotide chains to increase the stability of the double helix.

In the present invention, the "antisense" refers to a sequence of nucleotide bases in which an antisense oligomer is hybridized with a target sequence in RNA by Watson-Crick base pairing, allowing formation of typically mRNA and RNA: oligomeric heteroduplexes in the target sequence. and an oligomer having an inter-subunit backbone. Oligomers may have exact sequence complementarity or near complementarity to the target sequence.

Since the information of the AcAT protein according to the present invention or the gene encoding it is known, those skilled in the art can easily design primers, probes, or antisense nucleotides that specifically bind to the gene encoding the protein based on this information. .

According to another embodiment of the present invention, it relates to a kit for diagnosing anticancer drug resistance comprising the composition for diagnosing anticancer drug resistance of the present invention.

In the present invention, the "kit" refers to a tool capable of evaluating the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. Not only direct labeling of a detectable substance in relation to a probe or antibody by reaction with a substrate, but also indirect labeling in which a color-developing label is conjugated by reactivity with another directly labeled reagent. It may include a color-developing substrate solution, a washing solution, and other solutions that will undergo a color-reacting reaction with the marker, and may be prepared including reagent components to be used. In the present invention, the kit may be a kit containing essential elements necessary for performing RT-PCR, and in addition to each primer pair specific for the marker gene, a test tube, reaction buffer, deoxynucleotides (dNTPs), Taq-polymerization enzymes, reverse transcriptase, DNase, RNase inhibitors, sterile water, and the like. In addition, the kit may be a kit for detecting a gene for diagnosing anticancer drug resistance, including essential elements required to perform a DNA chip. The DNA chip kit includes a substrate to which a cDNA corresponding to a gene or a fragment thereof is attached as a probe, and the substrate may include a cDNA corresponding to a quantitative control gene or a fragment thereof. The kit of the present invention is not limited thereto, as long as it is known in the art.

In the present invention, the kit may be a RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or a multiple reaction monitoring (MRM) kit.

The kit of the present invention may further include one or more other component compositions, solutions or devices suitable for the assay method. For example, in the present invention, the kit may further include essential elements necessary for carrying out the reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit contains a pair of primers specific for a gene encoding a marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene, and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. In addition, a primer specific for the nucleic acid sequence of the control gene may be included. Other reverse transcription polymerase reaction kits contain a test tube or other suitable container, reaction buffer (with varying pH and magnesium concentration), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase and RNase inhibitor DEPC. -Water (DEPC-water), sterilized water, etc. may be included.

In addition, the kit for diagnosing anticancer drug resistance of the present invention may include essential elements required to perform a DNA chip. The DNA chip kit may include a substrate to which cDNA or oligonucleotides corresponding to genes or fragments thereof are attached, and reagents, reagents, enzymes, and the like for producing fluorescently labeled probes. In addition, the substrate may include a cDNA or oligonucleotide corresponding to a control gene or a fragment thereof.

In addition, the kit for diagnosing anticancer drug resistance of the present invention may include essential elements required to perform ELISA. ELISA kits contain antibodies specific for the protein. An antibody is an antibody that has high specificity and affinity for a marker protein and little cross-reactivity to other proteins, and is a monoclonal antibody, polyclonal antibody, or recombinant antibody. ELISA kits may also include antibodies specific for a control protein. Other ELISA kits include reagents capable of detecting bound antibodies, such as labeled secondary antibodies, chromophores, enzymes (eg, conjugated with antibodies) and substrates thereof or those capable of binding the antibody. may contain other substances and the like.

In the kit for diagnosing anticancer drug resistance of the present invention, the fixture for the antigen-antibody binding reaction includes a nitrocellulose membrane, a PVDF membrane, a well plate made of polyvinyl resin or polystyrene resin, and a well plate made of glass. A slide glass or the like may be used, but is not limited thereto.

In addition, in the kit for diagnosing anticancer drug resistance of the present invention, the marker of the secondary antibody is preferably a conventional colorant that reacts to color, and HRP (horseradish peroxidase), alkaline phosphatase, colloid gold, Labels such as fluorescein and dyes such as FITC (poly L-lysine-fluorescein isothiocyanate) and RITC (rhodamine-B-isothiocyanate) may be used, but are limited thereto It is not.

In addition, the chromogenic substrate for inducing color development in the kit for diagnosing anticancer drug resistance of the present invention is preferably used according to a marker that produces a color reaction, and TMB (3,3',5,5'-tetramethyl bezidine), ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)], OPD (o-phenylenediamine), and the like can be used. At this time, it is more preferable that the chromogenic substrate is provided in a state dissolved in a buffer solution (0.1 M NaAc, pH 5.5). A chromogenic substrate such as TMB is decomposed by HRP used as a marker for the secondary antibody conjugate to generate a chromogenic deposit, and the presence or absence of the marker proteins is detected by visually checking the degree of chromogenic deposit.

In the anticancer drug resistance diagnostic kit of the present invention, the washing solution preferably includes a phosphate buffer solution, NaCl, and Tween 20, and a buffer solution (PBST) composed of 0.02 M phosphate buffer solution, 0.13 M NaCl, and 0.05% Tween 20. ) is more preferred. The washing solution is washed 3 to 6 times by reacting the secondary antibody with the antigen-antibody conjugate after the antigen-antibody binding reaction, and then adding an appropriate amount to the stationary body. As the solution for stopping the reaction, a sulfuric acid solution (H2SO4) may be preferably used.

According to another embodiment of the present invention, it relates to a method for providing information for diagnosing anticancer drug resistance.

The method of the present invention may include measuring the expression level of the AcAT protein or a gene encoding the protein in a biological sample isolated from the target subject.

The method of the present invention may be for screening for the presence or absence of anticancer drug resistance in a biological sample isolated from a subject of interest.

In the present invention, the "object of interest" is an individual who has developed cancer or has a high possibility of developing cancer, and may be mammals including humans, such as humans, rats, mice, guinea pigs, hamsters, rabbits, monkeys, and dogs. , may be selected from the group consisting of cats, cows, horses, pigs, sheep and goats, and may be preferably human, but is not limited thereto.

In the present invention, the biological sample refers to any material, biological fluid, tissue, or cell obtained from or derived from an individual, including whole blood, leukocytes, and peripheral blood mononuclear cells. ), buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, respiration (breath), urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid , glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint aspirate (joint aspirate), organ secretions, cells (cell), cell extract (cell extract) and cerebrospinal fluid (cerebrospinal fluid) may be at least one selected from the group consisting of, but is not limited thereto.

In the present invention, the agent for measuring the expression level of the AcAT protein is at least one selected from the group consisting of an antibody, an oligopeptide, a ligand, a peptide nucleic acid (PNA), and an aptamer that binds specifically to the AcAT protein can include

In the present invention, the expression level of the AcAT protein is measured by protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption) /Ionization Time of Flight Mass Spectrometry) assay, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoretic assay, liquid chromatography-mass spectrometry (liquid chromatography-mass spectrometry, LC-MS), LC-MS/MS (liquid chromatography-mass spectrometry/mass spectrometry), western blotting, or ELISA (enzyme linked immunosorbentassay).

In addition, in the present invention, the expression level of the AcAT protein may be measured by a multiple reaction monitoring (MRM) method.

In the multiple reaction monitoring method in the present invention, a synthetic peptide or E. coli beta-galactosidase in which a specific amino acid constituting the target peptide is substituted with an isotope may be used as an internal standard material.

In the present invention, the AcAT protein may consist of the amino acid sequence represented by SEQ ID NO: 1, but is not limited thereto.

In the present invention, the agent for measuring the expression level of the gene encoding the AcAT protein may include at least one selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the gene encoding the AcAT protein. .

In the present invention, measurement of the expression level of the gene encoding the AcAT protein is reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction (Competitive RT-PCR), real-time reverse transcription polymerase reaction (Real-time RT-PCR) ), RNase protection assay (RPA), Northern blotting, or DNA chip.

In the information providing method of the present invention, descriptions of antibodies, oligopeptides, ligands, peptide nucleic acids (PNAs), aptamers, etc. and descriptions of primers, probes, antisense nucleotides, etc. are overlapped with those described above, thereby complicating the specification excessively. In order to avoid this, the detailed description is omitted below.

In the present invention, when the expression level of the AcAT protein or the gene encoding it measured for the biological sample of the target subject is higher than that of the control group, it is predicted that the target subject has developed or is likely to develop anticancer drug resistance, It can be predicted that the treatment response to anticancer drugs is low or that the prognosis of cancer treatment is poor.

In the present invention, the term "control group" may be a normal control group in which resistance to anticancer drugs does not occur, or an average to a median level of expression of the AcAT protein or a gene encoding the same in anticancer drug sensitive cells. The expression level of the marker protein or nucleic acid molecule encoding it in the control group and the expression level of the marker protein or nucleic acid molecule encoding it in the biological sample derived from the cancer patient to be analyzed can be compared, and whether or not there is a significant change in the expression level It is possible to diagnose anticancer drug resistance by determining.

In the present invention, the anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, cedi ranib, lestaurtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, bevacizumab, cisplatin, cetuximab, viscum album, asparaginase, tretinoin, hydroxycarbamide, Dasatinib, estramustine, gemtuzumab ozogamicin, ibritumomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, chitosan holmium nitrate, gemcitabine, doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacytidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludabine, enocy Tabine, flutamide, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, bin Blastin, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pirarubicin, aclarubicin, pepromycin, temsy lolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, x with mestanes, aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine, and carmustine It may be one using a drug containing at least one selected from the group consisting of, more preferably 5-fluorouracil or oxaliplatin, but is not limited thereto. The description of the drugs is duplicated with the previous description, and thus the detailed description thereof is omitted below to avoid excessive complexity of the specification.

In the present invention, the cancer is thyroid cancer, parathyroid cancer, gastric cancer, ovarian cancer, colon cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, and Hodgkin's lymphoma. , blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, perianal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine adenocarcinoma , adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteric cancer, renal cell carcinoma, renal pelvic carcinoma, CNS central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma , tumor differentiation and/or proliferation, etc., as long as the cancer progression is dependent on the cancer cells and/or cancer stem cells described in the present invention, it is not limited thereto.

2. Anticancer drug resistance treatment use

According to another embodiment of the present invention, it relates to a pharmaceutical composition for treating anticancer drug resistance or enhancing anticancer drug sensitivity.

The composition of the present invention is an agent that reduces the activity or expression level of AcAT protein; Alternatively, it may include an agent that reduces the expression level of the gene encoding the protein as an active ingredient.

The agent for reducing the activity or expression level of the protein of the present invention is any one selected from the group consisting of compounds, peptides, peptide mimetics, aptamers, antibodies, and natural products that specifically bind to the protein or a part thereof. It may include, but is not limited to, a method known as a method commonly used in the art that corresponds to a means for deriving an effect of inhibiting its activity or expression by directly or indirectly acting on the target AcAT protein. If it can be easily derived by technology, it is not limited thereto and may include all.

In the present invention, the "peptide mimetics" are peptides or non-peptides that inhibit the binding domain of AcAT protein leading to inhibition of AcAT activity. Key residues of non-hydrolysable peptide analogs include the β-turn dipeptide core (Nagai et al. Tetrahedron Lett 26:647, 1985), keto-methylene pseudopeptides (Ewenson et al. J Med chem 29:295, 1986; and Ewenson et al. in Peptides: Structure and Function (Proceedings of the 9th AmeriCan Peptide Symposium) Pierce chemiCal co. Rockland, IL, 1985), azepine (Huffman et al. in Peptides: chemistry and Biology, G.R. Marshall ed., EScOM Publisher: Leiden, Netherlands, 1988), benzodiazepines (Freidinger et al. in Peptides; chemistry and Biology, G.R. Marshall ed., EScOM Publisher: Leiden, Netherlands, 1988), β-aminoalcohols (Gordon et al. Biochem Biophys Res. commun 126:419 1985) and substituted gamma-lactam rings (Garvey et al. in Peptides: Chemistry and Biology, G.R. Marshell ed., EScOM Publisher: Leiden, Netherlands, 1988).

In the present invention, the "Aptamer" is a single-stranded nucleic acid (DNA, RNA or modified nucleic acid) that has a stable tertiary structure and can bind to a target molecule with high affinity and specificity. Since the aptamer discovery technology called SELEX (Systematic Evolution of Ligands by EXponential Enrichment) was first developed (Ellington, AD and Szostak, JW., Nature 346:818-822, 1990), aptamers have been developed from low-molecular-weight organic substances to peptides and membrane proteins. Many aptamers capable of binding to various target molecules have been continuously discovered. Aptamers are comparable to single antibodies because of their inherent high affinity (usually pM level) and specificity of being able to bind to target molecules, and in particular, they have high potential as alternative antibodies to the extent that they are called "chemical antibodies."

In the present invention, the "antibodies" prepared by injecting the protein or purchased commercially can be used. In addition, the antibody includes polyclonal antibodies, monoclonal antibodies, fragments capable of binding to an epitope, and the like.

Here, the polyclonal antibody can be produced by a conventional method of injecting the protein into an animal and collecting blood from the animal to obtain antibody-containing serum. Such polyclonal antibodies can be purified by any method known in the art, and can be made from any animal species host, such as goat, rabbit, sheep, monkey, horse, pig, cow, dog, etc.

In addition, the monoclonal antibody can be produced using any technique that provides for the production of antibody molecules through the cultivation of continuous cell lines. Such technologies include, but are not limited to, hybridoma technology, human B-cell line hybridoma technology, and EBV-hybridoma technology.

In addition, antibody fragments containing specific binding sites for the above proteins can be prepared. For example, but not limited to, F(ab')2 fragments can be prepared by pepsin digestion of antibody molecules, and Fab fragments can be prepared by reducing disulfide bridges of F(ab')2 fragments. Alternatively, by miniaturizing the Fab expression library, monoclonal Fab fragments having the desired specificity can be quickly and conveniently identified.

In the present invention, the antibody may be bound to a solid substrate to facilitate subsequent steps such as washing or separation of complexes. Solid substrates include, for example, synthetic resins, nitrocellulose, glass substrates, metal substrates, glass fibers, microspheres and microbeads. In addition, the synthetic resin includes polyester, polyvinyl chloride, polystyrene, polypropylene, PVDF, and nylon.

In the present invention, the agent that reduces the activity or expression level of the protein may be one that specifically binds to the polypeptide represented by SEQ ID NO: 1 of the AcAT protein, and preferably, the composition of the present invention is an antibody specific to the AcAT protein. It may include, wherein the antibody may specifically bind to the polypeptide represented by SEQ ID NO: 1, but is not limited thereto.

The agent for reducing the expression level of the gene encoding the protein of the present invention is a gene encoding the protein, preferably an antisense nucleotide that binds complementary to the gene or a part thereof, a small interfering RNA (short interfering RNA; siRNA), short hairpin RNA, and ribozyme, but is not limited thereto, and acts directly or indirectly on the gene encoding the target AcAT protein As long as it is a means for deriving an effect in which the expression is inhibited, and can be easily derived by a known technique as a method commonly used in the art, it is not limited thereto and may include all.

In the present invention, the "antisense nucleotide", as defined in Watson-Crick base pairing, binds (hybridizes) to a complementary nucleotide sequence of DNA, immature-mRNA or mature mRNA to interfere with the flow of genetic information from DNA to protein will be. The specific nature of antisense nucleotides for target sequences makes them exceptionally multifunctional. Since antisense nucleotides are long chains of monomeric units, they can be easily synthesized against the target RNA sequence. Recently, many studies have demonstrated the usefulness of antisense nucleotides as a biochemical means to study target proteins. Since many recent advances have been made in the field of oligonucleotide chemistry and the synthesis of nucleotides that exhibit improved cell line adsorption, target binding affinity, and nuclease resistance, the use of antisense nucleotides can be considered as a new type of inhibitor.

In the present invention, the "shRNA" and "siRNA" are nucleic acid molecules capable of mediating RNA interference or gene silencing, and can suppress the expression of a target gene, thereby providing an efficient gene knockdown method or gene therapy used in a way shRNA is a hairpin structure formed by binding between complementary sequences within a single-stranded oligonucleotide, and in vivo, the shRNA is cleaved by dicer to form small RNA fragments of 21 to 25 nucleotides in size siRNA, which is a double-stranded oligonucleotide, can specifically bind to mRNA with a complementary sequence and suppress its expression. In addition, siRNA refers to a small double-stranded RNA fragment of 21 to 25 nucleotides in size that induces RNA interference (RNAi) by modifying target mRNA by double-stranded RNA (dsRNA).

In the present invention, which means of the shRNA and siRNA to be used can be determined by a person skilled in the art, and a similar expression reduction effect can be expected if the mRNA sequences they target are the same. For the purpose of the present invention, it is possible to suppress the expression of AcAT by specifically acting on the gene encoding the aTAT1 protein to cleave the aTAT1 gene (eg, mRNA molecule) to induce RNA interference (RNAi). can siRNA can be synthesized chemically or enzymatically. The method for preparing siRNA is not particularly limited, and methods known in the art may be used. For example, a method for chemically synthesizing siRNA directly, a method for synthesizing siRNA using in vitro transcription, a method for cutting long double-stranded RNA synthesized by in vitro transcription using an enzyme, An expression method through intracellular delivery of shRNA expression plasmids or viral vectors and an expression method through intracellular delivery of polymerase chain reaction (PCR)-induced siRNA expression cassettes, but are not limited thereto.

In the present invention, the "ribozyme" refers to an RNA molecule having a catalytic activity. Ribozymes with various activities are known, the ribozymes of the AcAT gene include known or artificially produced ribozymes, and optionally ribozymes having target-specific RNA cleavage activity are prepared by known standard techniques It can be.

Agents that reduce the activity or expression of the AcAT (acetylated alpha tubulin) protein in the present invention; Alternatively, the agent reducing the expression level of the gene encoding the protein may be one that inhibits acetylation of the AcAT protein, for example, Histone Deacetylase 6 (HDAC6) protein, HDAC6 protein expression promoters or activators of HDAC6 protein; or alpha-tubulin N-acetyltransferase (aTAT1) protein activity or expression inhibitor or expression inhibitor of the gene encoding the aTAT1 protein, but acetylation of acetylated tubulin protein As long as it has the function of controlling anger, it is not limited thereto and may include all of them.

In the present invention, the "Histone Deacetylase 6 (HDAC6)" is an enzyme encoded by the human HDAC6 gene, and acetylation/deacetylation is involved in the angiogenesis process, and angiogenesis is induced under hypoxic conditions to treat cancer. The proliferation of cells is active, which is due to the increased activity of HDAC (histone deacetylase) under hypoxic conditions, which suppresses the action of tumor suppressors and induces angiogenesis. Histone deacetylases are histone-modifying enzymes that participate in transcriptional repression by removing acetyl groups from target histones. In the present invention, information about the HDAC6 protein or the gene encoding the same is registered in NCBI (National Center for Biotechnology Information), and the HDAC6 protein may consist of the amino acid sequence represented by SEQ ID NO: 3, but is not limited thereto, As non-limiting examples, at least 99% and less than 100%, at least 95% and less than 99%, at least 90% and less than 95%, at least 85% and less than 90%, or at least 80% and less than 85% the sequence of HDAC6. It may be the case of having the homology of, and it may include all without limitation within the obvious range that it exerts the desired effect of the present invention to those skilled in the art.

In the present invention, the "HDAC6 protein expression promoter" is a substance that enhances the expression of the HDAC6 protein, and includes all substances that enhance the expression of HDAC6 at the transcriptional level or protein level. For example, HDAC6 protein It may include, but is not limited to, coding genes, expression vectors containing the same, and the like.

In the present invention, the "activator of HDAC6 protein" means a substance that positively regulates the activity of HDAC6 protein by directly interacting with or indirectly interacting with HDAC6 protein. For example, the HDAC6 protein activator may be a substance that binds to the HDAC6 protein and enhances the activity of the HDAC6 protein.

In the present invention, the agent that reduces the expression level of the gene encoding the aTAT1 protein is an antisense nucleotide that binds complementary to the gene encoding the aTAT1 protein, small interfering RNA (siRNA), short hairpin RNA (short It may include one or more selected from the group consisting of hairpin RNA) and ribozyme, but is not limited thereto.

In the present invention, the "alpha tubulin N-acetyl transferase (aTAT1)" is an enzyme belonging to the transferase family encoded by the ATAT1 gene, and is also called TAT, C6orf134, Nbla00487, alpha-TAT, or alpha-TAT1. It belongs to the transferase family, and more specifically to acyl transferases. The ATAT1 gene encodes a clathrin protein that plays an important role in vesicle formation and acetylates alpha tubulin on lysine 40 (K40). This process is known to be important for microtubule growth during chemotaxis and cilium formation. In the present invention, information about the aTAT1 protein or the gene encoding the same is registered in NCBI (National Center for Biotechnology Information), and the aTAT1 protein may consist of the amino acid sequence represented by SEQ ID NO: 4, but is not limited thereto, As non-limiting examples, at least 99% and less than 100%, at least 95% and less than 99%, at least 90% and less than 95%, at least 85% and less than 90%, or at least 80% and less than 85% the sequence of HDAC6. It may be the case of having the homology of, and it may include all without limitation within the obvious range that it exerts the desired effect of the present invention to those skilled in the art.

As an example of the present invention, as an agent for reducing the expression level of the gene encoding the aTAT1 protein, SEQ ID NO: 5 and SEQ ID NO: 6; SEQ ID NO: 7 and SEQ ID NO: 8; or SEQ ID NO: 9 and SEQ ID NO: 10; may be a siRNA consisting of a base sequence represented by at least one combination selected from the group consisting of, but is not limited thereto.

The composition of the present invention, in order to treat anticancer drug resistance or increase the anticancer drug sensitivity enhancing effect, histone deacetylase 6 (Histone Deacetylase 6; HDAC6) protein, HDAC6 protein expression promoter or activator of HDAC6 protein; Alternatively, an alpha-tubulin N-acetyltransferase (aTAT1) protein activity or expression inhibitor or an expression inhibitor of a gene encoding the aTAT1 protein may be further included.

In addition, the composition of the present invention may further include an anticancer agent, wherein the anticancer agent includes nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, Nirotinib, Semasanib, Bosutinib, Axitinib, Cediranib, Lestaurtinib, Trastuzumab, Gefitinib, Bortezomib, Sunitinib, Carboplatin, Bevacizumab, Cisplatin, Cetuximab , viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine, gemtuzumabozogamicin, ibritumomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, al Remtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine, doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacitidine, methotrexate, uracil , cytarabine, 5-fluorouracil, fludabine, enocitabine, flutamide, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, velo Tecan, Topotecan, Vinorelbine, Etoposide, Vincristine, Vinblastine, Teniposide, Doxorubicin, Idarubicin, Epirubicin, Mitoxantrone, Mitomycin, Bleromycin, Daunorubicin, Dactino Mycin, pirarubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, chiotepa, nimustine , chlorambucil, mitolactol, leucovorin, tretonin, exmestane, aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, retro It may be a drug containing at least one selected from the group consisting of sol, vorozol, bicalutamide, lomustine and carmustine, and more preferably 5-fluorouracil or oxaliplatin, but is limited thereto it is not going to be

In the present invention, the "anti-cancer drug resistance" refers to the decrease in the effectiveness of the drug when the anti-cancer drug is used repeatedly in a quantitative manner, and in order to obtain the same effect previously experienced by patients with anti-cancer drug resistance, the amount of use or the frequency of use must be increased It is a condition in which the same effect cannot be obtained even if the substance needs to be increased or administered in the same amount as before.

In the present invention, the "resistant treatment" means that when the anticancer drug is used repeatedly in a quantitative manner, the effect of the drug is reduced, or the amount or frequency of use must be increased in order to obtain the same effect previously experienced in patients with anticancer drug resistance. It refers to the action of restoring a condition in which the same effect as before is not obtained even if the same amount of substance is administered. More specifically, the same anticancer effect can be obtained even when the anticancer drug is applied less frequently or at a lower dose, or the same effect can be obtained even if the same dose or a lower dose is administered by returning to the state before anticancer drug resistance occurs. It refers to the action of creating a state.

In the pharmaceutical composition for treating anticancer drug resistance and the composition for enhancing anticancer drug sensitivity of the present invention, descriptions of the AcAT protein or the gene encoding the same, various anticancer agents, etc. are the same as those described in the preceding diagnostic composition, in order to avoid excessive complexity in the present specification. omit

According to another embodiment of the present invention, it relates to a pharmaceutical composition for preventing or treating anticancer drug-resistant cancer.

The composition of the present invention is an agent that reduces the activity or expression level of AcAT protein; Alternatively, it may include an agent that reduces the expression level of the gene encoding the protein as an active ingredient.

Agents that reduce the activity or expression of the AcAT (acetylated alpha tubulin) protein in the present invention; Alternatively, the agent reducing the expression level of the gene encoding the protein may be one that inhibits acetylation of the AcAT protein, for example, Histone Deacetylase 6 (HDAC6) protein, HDAC6 protein expression promoters or activators of HDAC6 protein; or alpha-tubulin N-acetyltransferase (aTAT1) protein activity or expression inhibitor or expression inhibitor of the gene encoding the aTAT1 protein, but acetylation of acetylated tubulin protein As long as it has the function of controlling anger, it is not limited thereto and may include all of them.

The composition of the present invention, in order to treat anticancer drug resistance or increase the anticancer drug sensitivity enhancing effect, histone deacetylase 6 (Histone Deacetylase 6; HDAC6) protein, HDAC6 protein expression promoter or activator of HDAC6 protein; Alternatively, an alpha-tubulin N-acetyltransferase (aTAT1) protein activity or expression inhibitor or an expression inhibitor of a gene encoding the aTAT1 protein may be further included.

In addition, the composition of the present invention may further include an anticancer agent, wherein the anticancer agent includes nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, Nirotinib, Semasanib, Bosutinib, Axitinib, Cediranib, Lestaurtinib, Trastuzumab, Gefitinib, Bortezomib, Sunitinib, Carboplatin, Bevacizumab, Cisplatin, Cetuximab , viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine, gemtuzumabozogamicin, ibritumomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, al Remtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine, doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacitidine, methotrexate, uracil , cytarabine, 5-fluorouracil, fludabine, enocitabine, flutamide, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, velo Tecan, Topotecan, Vinorelbine, Etoposide, Vincristine, Vinblastine, Teniposide, Doxorubicin, Idarubicin, Epirubicin, Mitoxantrone, Mitomycin, Bleromycin, Daunorubicin, Dactino Mycin, pirarubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, chiotepa, nimustine , chlorambucil, mitolactol, leucovorin, tretonin, exmestane, aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, retro It may be a drug containing at least one selected from the group consisting of sol, vorozol, bicalutamide, lomustine and carmustine, and more preferably 5-fluorouracil or oxaliplatin, but is limited thereto it is not going to be

The "prevention" of the present invention may be included without limitation as long as it is an act of blocking, suppressing or delaying symptoms caused by uncontrolled growth of cancer cells by using the composition of the present invention.

The "treatment" of the present invention may be included without limitation as long as symptoms caused by uncontrolled growth of cancer cells are improved or beneficial by using the composition of the present invention.

In the pharmaceutical composition for preventing or treating anticancer drug-resistant cancer of the present invention, each protein or a gene encoding the same, an agent reducing the activity or expression level of the protein; Or, the description of an agent that reduces the expression level of the gene encoding the protein is the same as that described in the composition for treating anticancer drug resistance and enhancing anticancer drug sensitivity, and thus is omitted to avoid excessive complexity in the present specification.

In the present invention, the pharmaceutical composition may be in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be for humans.

The pharmaceutical compositions of the present invention are not limited thereto, but are formulated in the form of oral formulations such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories and sterile injection solutions, respectively, according to conventional methods. can be used The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavors, etc. for oral administration. In the case of injections, buffers, preservatives, painless A topical agent, a solubilizer, an isotonic agent, a stabilizer, and the like may be mixed and used, and in the case of topical administration, a base, an excipient, a lubricant, a preservative, and the like may be used. The dosage form of the pharmaceutical composition of the present invention may be variously prepared by mixing with a pharmaceutically acceptable carrier as described above. For example, for oral administration, it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and in the case of injections, it can be prepared in unit dosage ampoules or multiple dosage forms. there is. In addition, it may be formulated into solutions, suspensions, tablets, capsules, sustained-release preparations, and the like.

On the other hand, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil and the like can be used. In addition, fillers, anti-agglomerating agents, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, and the like may be further included.

The route of administration of the pharmaceutical composition of the present invention is not limited to oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical , sublingual or rectal. Oral or parenteral administration is preferred.

The "parenteral" of the present invention includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. Preferably, the pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration, but is not limited thereto.

The pharmaceutical composition of the present invention depends on various factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, route of administration, excretion rate, drug combination and severity of the specific disease to be prevented or treated. The dosage of the pharmaceutical composition varies depending on the patient's condition, body weight, disease severity, drug form, administration route and period, but may be appropriately selected by those skilled in the art, and is 0.0001 to 50 mg per day. /kg or 0.001 to 50 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The dosage is not intended to limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated into a pill, dragee, capsule, liquid, gel, syrup, slurry, or suspension.

3. Screening method for drugs for overcoming or treating anticancer drug resistance or for enhancing sensitivity to anticancer drugs

According to another embodiment of the present invention, it relates to a method for screening a drug for overcoming or treating anticancer drug resistance or a drug for enhancing sensitivity to an anticancer drug.

The screening method of the present invention includes the steps of in vitro processing a candidate substance for cancer cells or cancer tissues expressing an AcAT protein or a gene encoding the same; and measuring the activity or expression level of the AcAT protein or the expression level of a gene encoding the protein in the cancer cells or cancer tissues after treatment with the candidate substance.

In the present invention, the "screening" means to select a substance having a specific target property from a candidate group composed of various substances by a specific manipulation or evaluation method.

In the present invention, the cancer cells may be isolated from a target individual, preferably an individual resistant to or likely to be resistant to anticancer drugs, or may be engineered to overexpress the AcAT protein or a gene encoding the same, preferably A recombinant vector containing a gene encoding the AcAT protein may be introduced into cancer cells and transfected.

In the present invention, the "vector" means a means for expressing a target gene in a host cell. The vector includes elements for expression of a target gene, may include a replication origin, a promoter, an operator, a transcription termination sequence, and the like, and is incorporated into the genome of a host cell. A ribosome binding site (RBS) for translation into a protein and/or an appropriate enzyme site (e.g., restriction enzyme site) for incorporation and/or selectable markers to confirm successful incorporation into the host cell, IRES (Internal Ribosome Entry Site) and the like may be additionally included. The vector may be engineered by a conventional genetic engineering method to have the above fusion polynucleotide (fusion promoter) as a promoter. The vector may further include transcription control sequences (eg, enhancers, etc.) other than the promoter.

In the present invention, the recombinant vector may be a viral or non-viral vector, and the viral vector may be an adenovirus vector, a retroviral vector including lentivirus, an adeno-associated virus vector, or a herpes simplex virus vector. , but not limited thereto. In addition, the non-viral vector may be a plasmid vector, a bacteriophage vector, a liposome, a bacterial artificial chromosome, or a yeast artificial chromosome, but is not limited thereto.

In the present invention, the gene of interest in the recombinant vector may be operably linked to the fusion polynucleotide. The term “operably linked” refers to a functional linkage between a gene expression control sequence and another nucleotide sequence. The gene expression control sequences can be "operably linked" to control the transcription and/or translation of other nucleotide sequences. In the recombinant vector, in order for the fusion polynucleotide to be operably linked to the target gene, the fusion polynucleotide may be linked to the 5' end of the target gene. The recombinant vector of the present invention can be used as an expression vector for a target protein capable of expressing the target protein with high efficiency in an appropriate host cell when the gene encoding the target protein to be expressed is operably linked.

The recombinant vector of the present invention may further include a transcription control sequence. The transcription control sequence may include a transcription termination sequence such as a polyadenylation sequence (pA); It may be one or more species selected from the group consisting of replication origins such as f1 origin of replication, SV40 origin of replication, pMB1 origin of replication, adeno origin of replication, AAV origin of replication, BBV origin of replication, etc., but is not limited thereto.

In addition, in the present invention, the recombinant vector may further include a selectable marker. The selectable marker is a gene for confirming whether a recombinant vector has been successfully introduced into a host cell or for constructing a stable cell line, for example, one selected from the group consisting of a drug resistance gene such as antibiotics, a metabolism-related gene, and a gene amplification gene. may be ideal

In the present invention, delivery (introduction) of the recombinant vector into cancer cells may use a delivery method widely known in the art. The delivery method, for example, microinjection, calcium phosphate precipitation, electroporation, sonoporation, magnetofection using a magnetic field, liposome-mediated transfection, gene bombardment bombardment), the use of dendrimers and inorganic nanoparticles, etc. may be used, but is not limited thereto.

In the present invention, the candidate substance may be at least one selected from the group consisting of natural compounds, synthetic compounds, RNA, DNA, polypeptides, enzymes, proteins, ligands, antibodies, antigens, bacterial or fungal metabolites, and bioactive molecules. , but is not limited thereto.

In the present invention, the agent for measuring the activity or expression level of the above-described protein is not particularly limited, but examples thereof include antibodies, oligopeptides, ligands, PNA (peptide nucleic acid) and aptamers that specifically bind to the protein ) may include one or more selected from the group consisting of.

In the present invention, methods for measuring or comparing the activity or expression level of the protein include protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI- TOF (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoretic assay, Liquid chromatography-mass spectrometry (LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/ Mass Spectrometry), Western blotting or ELISA (enzyme linked immunosorbentassay), etc., but limited to these it is not going to be

In the present invention, the agent for measuring the expression level of the gene encoding the protein may include at least one selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the gene.

Since the information of the protein according to the present invention or the gene encoding it is known, those skilled in the art will be able to easily design primers, probes or antisense nucleotides specifically binding to the gene encoding the protein based on this information.

In the present invention, in the process of confirming the presence and expression level of the gene, analysis methods for measuring the expression level of the gene include reverse transcription polymerase reaction (RT-PCR) and competitive reverse transcription polymerase reaction (Competitive RT-PCR) , real-time reverse transcription polymerase reaction (Real-time RT-PCR), RNase protection assay (RPA; RNase protection assay), Northern blotting, or DNA chip, but is not limited thereto.

In the present invention, when the activity or expression level of the AcAT protein measured in the cancer cells or cancer tissues after treatment of the candidate material is reduced, or the expression level of the gene encoding the protein is decreased, the candidate material is resistant to anticancer drugs. A step of determining the drug as a drug for overcoming or treating or enhancing sensitivity to anticancer drugs may be further included.

In the screening method of the present invention, descriptions of anticancer drugs and cancer are the same as those described in the composition for diagnosing anticancer drug resistance, and thus are omitted to avoid excessive complexity in the present specification.

In the case of using the present invention, by diagnosing resistance to an anticancer drug, it is possible to provide accurate basic information on the suitability of an anticancer drug for each cancer patient in a future treatment plan. In addition, when the present invention is used, resistance to anticancer drugs can be overcome and furthermore, anticancer drug resistant cancers can be effectively prevented, improved or treated.

Figure 1a is a diagram confirming the AcAT gene expression level measured in gastric cancer cell lines SK4, MKN45, and AGS according to an embodiment of the present invention through Western blot analysis.
Figure 1b is a diagram confirming the cumulative survival rate for each group according to the level of AcAT expression through TMA analysis of 1015 gastric cancer patients according to an embodiment of the present invention.
2a to 2c show AcAT expression in parent cells (P cells) classified with gastric cancer stem cell surface markers (CD133 and CD44) and/or the degree of anticancer drug resistance according to an embodiment of the present invention by Western blotting and It is also confirmed through the MTS assay.
3a to 3c are IC50 values according to the degree of AcAT expression in P cells and selected cells (S cells) of gastric cancer cell lines SK4, MKN45, or AGS according to an embodiment of the present invention and the presence or absence of anticancer drug resistance. It is a diagram showing the amount of change.
4 is a diagram confirming AcAT expression and anticancer drug resistance in patient-derived organoid (PDO) according to an embodiment of the present invention.
Figure 5 is a diagram confirming the change in anticancer drug sensitivity according to the regulation of acetylation mechanism (Acetylation status) in SK4 cell line according to an embodiment of the present invention through Western blot.
6 is a diagram showing changes in anticancer drug sensitivity upon administration of an HDAC6 inhibitor drug according to an embodiment of the present invention.
7 is a diagram showing changes in sensitivity to anticancer drugs upon knockdown of aTAT1 (alpha tubulin acetyl transferase1) gene according to an embodiment of the present invention.
8 is a diagram confirming comparison of anticancer drug sensitivity upon administration of an acetylated tubulin target drug according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example 1: Confirmation of cumulative survival rate by group according to AcAT expression level

1.1 Preparation of anticancer drug resistant cells

The inventors of the present invention conducted all experiments with the approval of the Yonsei University College of Medicine Evaluation Board (Institutional Review Board, IRB). Human gastric cancer cell lines, SK4, MKN45, and AGS, were obtained from the Korean Cell Line Bank, and 1% penicillin-streptomycin (Gibco) was added to RPMI 1640 medium containing 10% FBS according to the guide of the Korean cell line bank. was added and cultured in a 37°C, 5% CO ₂ incubator (HERAcell 150i, Thermo Scientific, Waltham, MA, USA) and used for experiments.

1.2 AcAT gene expression level measured in gastric cancer cell lines SK4, MKN45, and AGS

The cell line obtained above was cultured into parent cells (parent cells; P cells) and parent cells were continuously cultured and divided into surviving cells (selected cells; S cells) after 30 days of Western blotting to confirm the level of protein expression of AcAT. performed. In all of the various gastric cancer cell lines, the expression level of the protein was relatively high in the case of S cells having stem cell characteristics (see FIG. 1a). A high level of AcAT expression suggests that the cure rate may be relatively low.

1.3 Cumulative survival rate by group according to AcAT expression level

This study was approved as a prospective study after deliberation by the Clinical Research Ethics Review Committee, and all patients were included in the study only if they gave their consent after hearing sufficient study explanations. 1015 gastric cancer patients were recruited, and paraffin-processed tissue sections were obtained from them and AcAT immunostaining was performed using tissue microarray (TMA) to analyze protein levels. It was confirmed that the cumulative survival rate in the patient group with high AcAT expression was lower than that in the patient group with relatively low AcAT expression (see FIG. 1b).

Example 2: Correlation between anticancer drug resistance and stem cell surface markers (CD133 and CD44)

Resistant gastric cancer cells were obtained by the method of Example 1, and the correlation between anticancer drug-resistant cells and stem cells was examined. Cancer stem cells are well known to have a lower anticancer treatment effect and a higher risk of recurrence than conventional cancer cells, and CD133 and CD44 correspond to indicators of cancer stem cells. The expression level of the AcAT gene in each group of parental cells (P cells) classified by the S cell group and the stem cell surface markers CD133 and CD44 was measured by Western blotting. It was confirmed that the results according to the presence or absence of the stem cell surface marker appeared in the mother cells in the same manner as in the S cells (see FIGS. 2a to 2c). More specifically, in the case of CD133-positive and CD44-positive parental cells, it can be seen that the expression level of AcAT is relatively higher than that of CD133-negative and CD44-negative parental cells. In addition, as a method of quantifying a drug response, for example, IC ₅₀ refers to the maximum concentration at the moment when the enzyme/protein activity of a cell drops by half when a drug is administered. Here, it was used as an indicator of cell activity. In the case of CD133 positive and CD44 positive, the expression level of AcAT is high, and in this case, the IC ₅₀ for 5-fluorouracil (5-Fu) and oxaliplatin is high, indicating anticancer drug resistance. The higher the IC ₅₀ concentration measured, the higher the resistance.

Example 3: Correlation between AcAT markers and anticancer drug resistance

Resistant gastric cancer cells were obtained by the method of Example 1, and the expression level of AcAT and IC ₅₀ concentrations for 5-fluorouracil (5-Fu) and oxaliplatin were measured in P cells and S cells, As shown in Figures 3a to 3c, it was confirmed that the expression level of AcAT significantly increased in S cells, and the IC of 5-fluorouracil (5-Fu) and oxaliplatin in S cells in which AcAT is highly expressed It was confirmed that anticancer drug resistance occurred from the fact that the concentration of ₅₀ was high.

When AcAT is highly expressed, resistance to 5-fluorouracil (5-Fu) or oxaliplatin (Oxaliplatin) drugs can be confirmed through a correlation between the marker and resistance development (see FIG. 4). In addition, the present inventors prepared patient-derived organoids (PDO) to observe changes in the level of markers in tissues, and as in the above example, the protein expression level was confirmed through Western blotting, and anticancer agents To see resistance, cell viability was analyzed through MTS assay. Cancer spheroids were prepared by the following method by supplementing growth factors using a serum-free culture medium. After transferring the cultured spheroids to a 15ml conical tube, filled with low-temperature PBS to 15ml, centrifuged at 1200 rpm for 5 minutes, and washing with PBS was repeated three times to remove the matrigel. Cells were fixed with 4% (w/v) paraformaldehyde (PFA), incubated for 30 minutes, and washed three times with PBS. Triton X-100 solution (in PBS; concentration: 1% (v/v)) was added to the cell sample and incubated for 1 hour, followed by Triton X-100 solution (in PBS; concentration: 2%). (v/v)) was added and washed twice. Then, a blocking buffer (BSA 0.279g, goat serum 450ul, Triton X-100 180ul, PBS 20X 450ul, DW 7920ul) was added and incubated for 30 minutes. BubR1 antibody (BD bioscience) was treated and incubated for 16 hours. Washed 3 times for 20 minutes with 0.2% (v/v) PBS-T (Triton X-100 solution in PBS). An HRP-tagged secondary antibody (Thermo Fisher Scientific) was added and incubated for 16 hours. Washed three times for 20 minutes with 0.2% (v/v) PBS-T, treated with FITC-conjugated tubulin antibody (Abcam) at a ratio of 1:1000, and incubated for 2 days. After washing three times for 20 minutes with 0.2% (v/v) PBS-T, DAPI was added and incubated for one day.

As a result of the above experiments, there was a difference in drug concentration that changes cell activity, and these differences are shown in FIG. 4 . The figure is a result of a 3D model system (PDO) using gastric cancer patient tissue, where N means an organoid made of normal tissue, T is an organoid made of tumor tissue, and each number represents a surgical patient means the number (case no.) of Referring to FIG. 4, the 293N and 316N groups were normal organoids, so the expression of AcAT was low, and the 200T, 215T, and 247T groups showed relatively high levels of AcAT. At this time, it was confirmed that a relatively low level of AcAT was measured in the 247T group among the tumor organoids, thereby further confirming the drug resistance of the two groups, 215T and 247T. As a result, resistance to 5-fluorouracil (5-Fu) and oxaliplatin, that is, resistance, was measured to be high in the 215T group in which AcAT was expressed at the highest level, and in the 247T group in which AcAT expression was relatively low, the above It was confirmed that the sensitivity to the drug was good. Since many anti-cancer drugs that are commonly developed have a certain degree of toxicity, if the same effect can be obtained even with low concentrations of drug administration, it can greatly contribute to the therapeutic effect.

Example 4: Confirmation of the possibility of anticancer drug resistance treatment through the regulation of acetylation mechanism

P cells and S cells were prepared from the SK4 gastric cancer cell line according to Example 1.1, P cells as a control group (PSK4-WT) and acetylation promoting group (PSK4-K40Q), S cells as a control group (SSK4-WT) and acetylation The results of measuring the expression level of the AcAT marker by Western blot were shown in FIG. 5 .

Referring to Figure 5, after administering 5-fluorouracil (5-Fu) and oxaliplatin drugs to each group, the change in IC ₅₀ concentration was confirmed, and it was digitized and displayed as a graph. As a result, the SSK4 control group In comparison, it was confirmed that anticancer drug resistance was lowered in the SSK4 group in which acetylation of lysine 40 was suppressed, and through this, it can be seen that anticancer drug resistance was overcome. On the other hand, it was confirmed that anticancer drug resistance in the PSK4 group in which acetylation of lysine 40 was promoted increased 4- to 5-fold or more compared to the PSK4 control group. This suggests that resistance to the drug can be treated by regulating the acetylation of K40. When the above results are put together and arranged in order of highest anticancer drug sensitivity, the PSK4-WT group, SSK4-K40R group, PSK4-K40Q group, and SSK4-WT group have good anticancer effects, while the PSK4-WT and SSK4-K40R groups have good anticancer effects. , PSK4-K40Q group and SSK4-WT group showed low anticancer effect.

Example 5: Confirmation of the possibility of anticancer drug resistance treatment through the regulation of HDAC and aTAT1 expression

In order to confirm the therapeutic effect through the regulation of the acetylation mechanism, the PSK4 and SSK4 cell groups prepared in Example 4 were treated with HDAC6 inhibitor drugs or aTAT1 expression was knocked down using siRNA, and the degree of anticancer drug resistance was measured by MTS assay. analyzed through At this time. The siRNAs used are shown in Table 1 below.

primer direction Sequence (5'→3') sequence number si aTAT1 #1 sense GAGAAGCACACUCUUGCUU SEQ ID NO: 5 antisense AAGCAAGAGUGUGCUUCUC SEQ ID NO: 6 si aTAT1 #2 sense GAGAGUAGAUCCAGAGUGU SEQ ID NO: 7 antisense ACACUCUGGAUCUACUCUC SEQ ID NO: 8 si aTAT1 #3 sense CAGUCCCACAGGUGAACAA SEQ ID NO: 9 antisense UUGUUCACCUGUGGGACUG SEQ ID NO: 10

Among the si aTAT1 list used in Table 1, the degree of anticancer drug resistance was further confirmed using si aTAT1 of #3, which was confirmed to have relatively effective knockdown (see FIG. 7). Experimental results Referring to FIGS. 6 and 7, when HDAC6 was inhibited, resistance to 5-fluorouracil (5-Fu) and oxaliplatin, that is, resistance to the drug, was increased in the PSK4 group. On the other hand, when looking at anticancer drug resistance when aTAT1 expression was knocked down using siRNA (#3), it was confirmed that resistance to the drug was lowered in the SSK4 group and the resistance was cured. On the other hand, YH21931 and Frost-450 among other anticancer drugs other than the above drugs were administered and analyzed through the MTS assay. Here, the YH21931 drug is the same drug as YH16899 (Kim DG et al., Nat Chem Biol. 2014 Jan; 10(1):29-34), and in addition to inhibition of metabolism, G2/M phase cell cycle arrest, caspase Frost-450 is a drug that targets microtubules and is a small molecule that inhibits microtubule polymerization and is used for various cancer treatments.

As a result of the experiment, it was confirmed that the P cell group with low AcAT expression and the S cell group with high AcAT expression showed the opposite results to those seen in the drugs 5-Fu and Oxaliplatin (FIG. 8). reference). This suggests that the treatment method for anticancer drug resistance depending on the type of drug may vary depending on the type of drug, and when the acetylation inhibitor (HDAC6 promoter or aTAT1 inhibitor) is administered in combination with other anticancer drugs, anticancer drug resistance is treated, resulting in anticancer activity. Synergistic effects can be induced.

Considering the above Examples 1 to 4 comprehensively, the AcAT marker can be utilized for the purpose of diagnosing anticancer drug resistance, and furthermore, it is possible to overcome 5-Fu and oxaliplatin drug resistance by promoting the expression of HDAC or inhibiting the expression of aTAT1. Therefore, it is expected that the anticancer effect can be remarkably improved by treating drug resistance by selecting patients with resistance after treatment with the corresponding drug.

Having described specific parts of the present invention in detail above, it is clear that these specific techniques are only preferred embodiments for those skilled in the art, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Composition for diagnosing or treating drug resistant cancer <130> PDPB201840k01 <150> KR 10-2020-0029938 <151> 2020-03-11 <160> 10 <170> KoPatentIn 3.0 <210> 1 <211> 451 <212> PRT <213> Homo sapiens <400> 1 Met Arg Glu Cys Ile Ser Ile His Val Gly Gln Ala Gly Val Gln Ile 1 5 10 15 Gly Asn Ala Cys Trp Glu Leu Tyr Cys Leu Glu His Gly Ile Gln Pro 20 25 30 Asp Gly Gln Met Pro Ser Asp Lys Thr Ile Gly Gly Gly Asp Asp Ser 35 40 45 Phe Asn Thr Phe Phe Ser Glu Thr Gly Ala Gly Lys His Val Pro Arg 50 55 60 Ala Val Phe Val Asp Leu Glu Pro Thr Val Ile Asp Glu Val Arg Thr 65 70 75 80 Gly Thr Tyr Arg Gln Leu Phe His Pro Glu Gln Leu Ile Thr Gly Lys 85 90 95 Glu Asp Ala Ala Asn Asn Tyr Ala Arg Gly His Tyr Thr Ile Gly Lys 100 105 110 Glu Ile Ile Asp Leu Val Leu Asp Arg Ile Arg Lys Leu Ala Asp Gln 115 120 125 Cys Thr Gly Leu Gln Gly Phe Leu Val Phe His Ser Phe Gly Gly Gly 130 135 140 Thr Gly Ser Gly Phe Thr Ser Leu Leu Met Glu Arg Leu Ser Val Asp 145 150 155 160 Tyr Gly Lys Lys Ser Lys Leu Glu Phe Ser Ile Tyr Pro Ala Pro Gln 165 170 175 Val Ser Thr Ala Val Val Glu Pro Tyr Asn Ser Ile Leu Thr Thr Thr His 180 185 190 Thr Thr Leu Glu His Ser Asp Cys Ala Phe Met Val Asp Asn Glu Ala 195 200 205 Ile Tyr Asp Ile Cys Arg Arg Asn Leu Asp Ile Glu Arg Pro Thr Tyr 210 215 220 Thr Asn Leu Asn Arg Leu Ile Gly Gln Ile Val Ser Ser Ile Thr Ala 225 230 235 240 Ser Leu Arg Phe Asp Gly Ala Leu Asn Val Asp Leu Thr Glu Phe Gln 245 250 255 Thr Asn Leu Val Pro Tyr Pro Arg Ile His Phe Pro Leu Ala Thr Tyr 260 265 270 Ala Pro Val Ile Ser Ala Glu Lys Ala Tyr His Glu Gln Leu Ser Val 275 280 285 Ala Glu Ile Thr Asn Ala Cys Phe Glu Pro Ala Asn Gln Met Val Lys 290 295 300 Cys Asp Pro Arg His Gly Lys Tyr Met Ala Cys Cys Leu Leu Tyr Arg 305 310 315 320 Gly Asp Val Val Pro Lys Asp Val Asn Ala Ala Ile Ala Thr Ile Lys 325 330 335 Thr Lys Arg Thr Ile Gln Phe Val Asp Trp Cys Pro Thr Gly Phe Lys 340 345 350 Val Gly Ile Asn Tyr Gln Pro Pro Thr Val Val Pro Gly Gly Asp Leu 355 360 365 Ala Lys Val Gln Arg Ala Val Cys Met Leu Ser Asn Thr Thr Ala Ile 370 375 380 Ala Glu Ala Trp Ala Arg Leu Asp His Lys Phe Asp Leu Met Tyr Ala 385 390 395 400 Lys Arg Ala Phe Val His Trp Tyr Val Gly Glu Gly Met Glu Glu Gly 405 410 415 Glu Phe Ser Glu Ala Arg Glu Asp Met Ala Ala Leu Glu Lys Asp Tyr 420 425 430 Glu Glu Val Gly Val Asp Ser Val Glu Gly Glu Gly Glu Glu Glu Glu Gly 435 440 445 Glu Glu Tyr 450 <210> 2 <211> 1356 <212> DNA <213 > Homo sapiens <400> 2 atgcgtgagt gcatctccat ccacgttggc caggctggtg tccagattgg caatgcctgc 60 tgggagctct actgcctgga acacggcatc cagcccgatg gccagatgcc aagtgacaag 120 accattgggg gaggagatga ttccttcaac accttcttca gtgagacggg ggctggcaag 180 catgtgcccc gggcagtgtt tgtagacttg gaacccacag tcattgatga agttcgcact 240 ggcacctacc gccagctctt ccaccctgag caacttatca caggcaaaga agatgctgcc 300 aataactatg cccgagggca ctacaccatt ggcaaggaga tcattgacct cgtgttggac 360 cgaattcgca agctggccga ccagtgcacg ggtctccagg gcttcttggt tttccacagc 420 tttggtgggg gaactggttc tgggttcacc tcgctgctca tggaacgtct ctcagttgat 480 tatggcaaga agtccaagct ggagttctct atttacccgg cgccccaggt ttccacagct 540 gtagttgagc cctacaactc catcctcacc acccacacca ccctggagca ctctgattgt 600 gccttcatgg tagacaatga ggccatctat gaca tctgtc gtagaaacct cgatattgag 660 cgtccaacct atactaacct gaataggtta ataggtcaaa ttgtgtcctc catcactgct 720 tccctgagat ttgatggagc cctgaatgtt gacctgacag aattccagac caacctggtg 780 ccctatcccc gcatccactt ccctctggcc acatatgccc ctgtcatctc tgctgagaaa 840 gcctaccatg aacagctttc tgtagcagag atcaccaatg cttgctttga gccagccaac 900 cagatggtga aatgtgaccc tcgccatggt aaatacatgg cttgctgcct gttgtaccgt 960 ggtgacgtgg ttcccaaaga tgtcaatgct gccattgcca ccatcaagac caagcgtacc 1020 atccagtttg tggattggtg ccccactggc ttcaaggttg gcatcaacta ccagcctccc 1080 actgtggtgc ctggtggaga cctggccaag gtacagagag ctgtgtgcat gctgagcaac 1140 accacagcca ttgctgaggc ctgggctcgc ctggaccaca agtttgacct gatgtatgcc 1200 aaacgtgcct ttgttcactg gtacgttggg gaggggatgg aggaaggtga gttttcagag 1260 gcccgtgagg acatggctgc ccttgagaag gattatgagg aggttggtgt ggattctgtt 1320 gaaggagagg gtgaggaaga aggagaggaa tactaa 1356 <210> 3 <211> 1215 <212> PRT <213 > Homo sapiens <400> 3 Met Thr Ser Thr Gly Gln Asp Ser Thr Thr Thr Arg Gln Arg Arg Ser 1 5 10 15 Ar g Gln Asn Pro Gln Ser Pro Pro Gln Asp Ser Ser Val Thr Ser Lys 20 25 30 Arg Asn Ile Lys Lys Gly Ala Val Pro Arg Ser Ile Pro Asn Leu Ala 35 40 45 Glu Val Lys Lys Lys Gly Lys Met Lys Lys Leu Gly Gln Ala Met Glu 50 55 60 Glu Asp Leu Ile Val Gly Leu Gln Gly Met Asp Leu Asn Leu Glu Ala 65 70 75 80 Glu Ala Leu Ala Gly Thr Gly Leu Val Leu Asp Glu Gln Leu Asn Glu 85 90 95 Phe His Cys Leu Trp Asp Asp Ser Phe Pro Glu Gly Pro Glu Arg Leu 100 105 110 His Ala Ile Lys Glu Gln Leu Ile Gln Glu Gly Leu Leu Asp Arg Cys 115 120 125 Val Ser Phe Gln Ala Arg Phe Ala Glu Lys Glu Glu Leu Met Leu Val 130 135 140 His Ser Leu Glu Tyr Ile Asp Leu Met Glu Thr Thr Gln Tyr Met Asn 145 150 155 160 Glu Gly Glu Leu Arg Val Leu Ala Asp Thr Tyr Asp Ser Val Tyr Leu 165 170 175 His Pro Asn Ser Tyr Ser Cys Ala Cys Leu Ala Ser Gly Ser Val Leu 180 185 190 Arg Leu Val Asp Ala Val Leu Gly Ala Glu Ile Arg Asn Gly Met Ala 195 200 205 Ile Ile Arg Pro Pro Gly His His Ala Gln His Ser Leu Met Asp Gly 210 215 220 Tyr Cys Met Phe Asn His Val Ala Val Ala Ala Arg Tyr Ala Gln Gln 225 230 235 240 Lys His Arg Ile Arg Arg Val Leu Ile Val Asp Trp Asp Val His 245 250 255 Gly Gln Gly Thr Gln Phe Thr Phe Asp Gln Asp Pro Ser Val Leu Tyr 260 265 270 Phe Ser Ile His Arg Tyr Glu Gln Gly Arg Phe Trp Pro His Leu Lys 275 280 285 Ala Ser Asn Trp Ser Thr Thr Gly Phe Gly Gln Gly Gln Gly Tyr Thr 290 295 300 Ile Asn Val Pro Trp Asn Gln Val Gly Met Arg Asp Ala Asp Tyr Ile 305 310 315 320 Ala Ala Phe Leu His Val Leu Leu Pro Val Ala Leu Glu Phe Gln Pr o 325 330 335 Gln Leu Val Leu Val Ala Ala Gly Phe Asp Ala Leu Gln Gly Asp Pro 340 345 350 Lys Gly Glu Met Ala Ala Thr Pro Ala Gly Phe Ala Gln Leu Thr His 355 360 365 Leu Leu Met Gly Leu Ala Gly Gly Lys Leu Ile Leu Ser Leu Glu Gly 370 375 380 Gly Tyr Asn Leu Arg Ala Leu Ala Glu Gly Val Ser Ala Ser Leu His 385 390 395 400 Thr Leu Leu Gly Asp Pro Cys Pro Met Leu Glu Ser Pro Gly Ala Pro 405 410 415 Cys Arg Ser Ala Gln Ala Ser Val Ser Cys Ala Leu Glu Ala Leu Glu 420 425 430 Pro Phe Trp Glu Val Leu Val Arg Ser Thr Glu Thr Val Glu Arg Asp 435 440 445 Asn Met Glu Glu Asp Asn Val Glu Glu Ser Glu Glu Glu Gly Pro Trp 450 455 460 Glu Pro Pro Val Leu Pro Ile Leu Thr Trp Pro Val Leu Gln Ser Arg 465 470 475 480 Thr Gly Leu Val Tyr Asp Gln Asn Met Met Asn His Cys Asn Leu Trp 485 490 495 Asp Ser His His Pro Glu Val Pro Gln Arg Ile Leu Arg Ile Met Cys 500 505 510 Arg Leu Glu Glu Leu Gly Leu Ala Gly Arg Cys Leu Thr Leu Thr Pro 515 520 525 Arg Pro Ala Thr Glu Ala Glu Leu Leu Thr Cys His Ser Ala Glu Tyr 530 535 540 Val Gly His Leu Arg Ala Thr Glu Lys Met Lys Thr Arg Glu Leu His 545 550 555 560 Arg Glu Ser Ser Asn Phe Asp Ser Ile Tyr Ile Cys Pro Ser Thr Phe 565 570 575 Ala Cys Ala Gln Leu Ala Thr Gly Ala Ala Cys Arg Leu Val Glu Ala 580 585 590 Val Leu Ser Gly Glu Val Leu Asn Gly Ala Ala Val Val Arg Pro Pro 595 600 605 Gly His His Ala Glu Gln Asp Ala Ala Cys Gly Phe Cys Phe Ph e Asn 610 615 620 Ser Val Ala Val Ala Ala Arg His Ala Gln Thr Ile Ser Gly His Ala 625 630 635 640 Leu Arg Ile Leu Ile Val Asp Trp Asp Val His Gly Asn Gly Thr 645 650 655 Gln His Met Phe Glu Asp Asp Pro Ser Val Leu Tyr Val Ser Leu His 660 665 670 Arg Tyr Asp His Gly Thr Phe Phe Pro Met Gly Asp Glu Gly Ala Ser 675 680 685 Ser Gln Ile Gly Arg Ala Ala Gly Thr Gly Phe Thr Val Asn Val Ala 690 695 700 Trp Asn Gly Pro Arg Met Gly Asp Ala Asp Tyr Leu Ala Ala Trp His 705 710 715 720 Arg Leu Val Leu Pro Ile Ala Tyr Glu Phe Asn Pro Glu Leu Val Leu 725 730 735 Val Ser Ala Gly Phe Asp Ala Ala Arg Gly Asp Pro Leu Gly Gly Cys 740 745 750 Gln Val Ser Pro Glu Gly Tyr Ala His Leu Thr Hi s Leu Leu Met Gly 755 760 765 Leu Ala Ser Gly Arg Ile Ile Leu Ile Leu Glu Gly Gly Tyr Asn Leu 770 775 780 Thr Ser Ile Ser Glu Ser Met Ala Ala Cys Thr Arg Ser Leu Leu Gly 785 790 795 800 Asp Pro Pro Pro Leu Leu Thr Leu Pro Arg Pro Pro Leu Ser Gly Ala 805 810 815 Leu Ala Ser Ile Thr Glu Thr Ile Gln Val His Arg Arg Tyr Trp Arg 820 825 830 Ser Leu Arg Val Met Lys Val Glu Asp Arg Glu Gly Pro Ser Ser Ser 835 840 845 Lys Leu Val Thr Lys Lys Ala Pro Gln Pro Ala Lys Pro Arg Leu Ala 850 855 860 Glu Arg Met Thr Thr Arg Glu Lys Lys Val Leu Glu Ala Gly Met Gly 865 870 875 880 Lys Val Thr Ser Ala Ser Phe Gly Glu Glu Ser Thr Pro Gly Gln Thr 885 890 895 Asn Ser Glu Thr Ala Val Val Ala Le u Thr Gln Asp Gln Pro Ser Glu 900 905 910 Ala Ala Thr Gly Gly Ala Thr Leu Ala Gln Thr Ile Ser Glu Ala Ala 915 920 925 Ile Gly Gly Ala Met Leu Gly Gln Thr Thr Ser Glu Glu Ala Val Gly 930 935 940 Gly Ala Thr Pro Asp Gln Thr Thr Ser Glu Glu Thr Val Gly Gly Ala 945 950 955 960 Ile Leu Asp Gln Thr Thr Ser Glu Asp Ala Val Gly Gly Ala Thr Leu 965 970 975 Gly Gln Thr Thr Ser Glu Ala Val Gly Gly Ala Thr Leu Ala Gln 980 985 990 Thr Thr Ser Glu Ala Ala Met Glu Gly Ala Thr Leu Asp Gln Thr Thr 995 1000 1005 Ser Glu Glu Ala Pro Gly Gly Thr Glu Leu Ile Gln Thr Pro Leu Ala 1010 1015 1020 Ser Ser Thr Asp His Gln Thr Pro Pro Thr Ser Pro Val Gln Gly Thr 1025 1030 1035 1040 Thr Pro Gln Ile Ser Pro Ser Thr Leu Ile Gly Ser Leu Arg Thr Leu 1045 1050 1055 Glu Leu Gly Ser Glu Ser Gln Gly Ala Ser Glu Se r Gln Ala Pro Gly 1060 1065 1070 Glu Glu Asn Leu Leu Gly Glu Ala Ala Gly Gly Gln Asp Met Ala Asp 1075 1080 1085 Ser Met Leu Met Gln Gly Ser Arg Gly Leu Thr Asp Gln Ala Ile Phe 1090 1095 1100 Tyr Ala Val Thr Pro Leu Pro Trp Cys Pro His Leu Val Ala Val Cys 1105 1110 1115 1120 Pro Ile Pro Ala Ala Gly Leu Asp Val Thr Gln Pro Cys Gly Asp Cys 1125 1130 1135 Gly Thr Ile Gln Glu Asn Trp Val Cys Leu Ser Cys Tyr Gln Val Tyr 1140 1145 1150 Cys Gly Arg Tyr Ile Asn Gly His Met Leu Gln His His Gly Asn Ser 1155 1160 1165 Gly His Pro Leu Val Leu Ser Tyr Ile Asp Leu Ser Ala Trp Cys Tyr 1170 1175 1180 Tyr Cys Gln Ala Tyr Val His His Gln Ala Leu Leu Asp Val Lys Asp 1185 1190 1195 1200 Ile Ala His Gln Asn Lys Phe Gly Glu Asp Met Pro His Pro His 1205 1210 1215 <210> 4 <211> 409 <212> PRT <213> Homo sapiens <400> 4 Met Trp Leu Thr Trp Pro Phe Cys Phe Leu Thr Ile Thr Leu Arg Glu 1 5 10 15 Glu Gly Val Cys His Leu Glu Ser Val Asp Leu Gln Gln Gln Ile Met 20 25 30 Thr Ile Ile Asp Glu Leu Gly Lys Al a Ser Ala Lys Ala Gln Asn Leu 35 40 45 Ser Ala Pro Ile Thr Ser Ala Ser Arg Met Gln Ser Asn Arg His Val 50 55 60 Val Tyr Ile Leu Lys Asp Ser Ser Ser Ala Arg Pro Ala Gly Lys Gly Ala 65 70 75 80 Ile Ile Gly Phe Ile Lys Val Gly Tyr Lys Lys Leu Phe Val Leu Asp 85 90 95 Asp Arg Glu Ala His Asn Glu Val Glu Pro Leu Cys Ile Leu Asp Phe 100 105 110 Tyr Ile His Glu Ser Val Gln Arg His Gly His Gly Arg Glu Leu Phe 115 120 125 Gln Tyr Met Leu Gln Lys Glu Arg Val Glu Pro His Gln Leu Ala Ile 130 135 140 Asp Arg Pro Ser Gln Lys Leu Leu Lys Phe Leu Asn Lys His Tyr Asn 145 150 155 160 Leu Glu Thr Thr Val Pro Gln Val Asn Asn Phe Val Ile Phe Glu Gly 165 170 175 Phe Phe Ala His Gln His Arg Pro Pro Ala Pro Ser Leu Arg Ala Thr 180 185 190 Arg His Ser Arg Ala Ala Ala Val Asp Pro Thr Pro Ala Ala Pro Ala 195 200 205 Arg Lys Leu Pro Pro Lys Arg Ala Glu Gly Asp Ile Lys Pro Tyr Ser 210 215 220 Ser Ser Asp Arg Glu Phe Leu Lys Val Ala Val Glu Pro Pro Trp Pro 225 230 235 240 Leu Asn Arg Ala Pro Arg Arg Ala Thr Pro Pro Ala His Pro Pro Pro 245 250 255 Arg Ser Ser Ser Leu Gly Asn Ser Pro Glu Arg Gly Pro Leu Arg Pro 260 265 270 Phe Val Pro Glu Gln Glu Leu Leu Arg Ser Leu Arg Leu Cys Pro Pro 275 280 285 His Pro Thr Ala Arg Leu Leu Leu Ala Ala Asp Pro Gly Gly Ser Pro 290 295 300 Ala Gln Arg Arg Arg Thr Arg Gly Thr Pro Pro Gly Leu Val Ala Gln 305 310 315 320 Ser Cys Cys Tyr Ser Arg His Gly Gly Val Asn Ser Ser Ser Pro Asn 325 330 335 Thr Gly Asn Gln Asp Ser Lys Gln Gly Glu Gln Glu Thr Lys Asn Arg 340 345 350 Ser Ala Ser Glu Glu Gln Ala Leu Ser Gln Asp Gly Ser Gly Glu Lys 355 360 365 Pro Met His Thr Ala Pro Pro Gln Ala Pro Ala Pro Pro Ala Gln Ser 370 375 380 Trp Thr Val Gly Gly Asp Ile Leu Asn Ala Arg Phe Ile Arg Asn Leu 385 390 395 400 Gln Glu Arg Arg Ser Thr Arg Pro Trp 405 <210> 5 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> sense si aTAT1 #1 <400> 5 gagaagcaca cucuugcuu 19 <210> 6 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> anti-sense si aTAT1 #1 <400> 6 aagcaagagu gugcuucuc 19 <210> 7 <211 > 19 <212> RNA <213> Artificial Sequence <220> <223> sense si aTAT1 #2 <400> 7 gagaguagau ccagagugu 19 <210> 8 <211> 19 <212> RNA <213> Artificial Sequence <220> < 223> anti-sense si aTAT1 #2 <400> 8 acacucugga ucuacucuc 19 <210> 9 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> sense si aTAT1 #3 <400> 9 cagucc caca ggugaacaa 19 <210> 10 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> anti-sense si aTAT1 #3<400> 10 uuguucaccu gugggacug 19

Claims (23)

A composition for diagnosing anticancer drug resistance in gastric cancer patients, comprising an agent for measuring the expression level of AcAT (acetylated alpha tubulin) protein. According to claim 1,
The agent for measuring the expression level of the protein comprises at least one selected from the group consisting of antibodies, oligopeptides, ligands, PNAs (peptide nucleic acids) and aptamers that bind specifically to the protein, composition . delete According to claim 1,
The anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, cediranib, lesta urtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, bevacizumab, cisplatin, cetuximab, viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib , estramustine, gemtuzumab ozogamicin, ibritumomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine, Doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacytidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludabine, enocitabine, flu Tamid, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pirarubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, exmestane, in the group consisting of aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine and carmustine A composition comprising at least one selected species. According to claim 4,
Wherein the anti-cancer agent is an antimetabolic agent or an alkylating agent. According to claim 5,
Wherein the antimetabolite drug is 5-Fluorouracil (5-FU), and the alkylating agent drug is oxaliplatin. According to claim 1,
The anticancer agent is for the treatment of gastric cancer, the composition. A kit for diagnosing anticancer drug resistance in gastric cancer patients, comprising the composition of any one of claims 1, 2, and 4 to 7. A method for providing information for diagnosing anticancer drug resistance in gastric cancer patients, comprising measuring the expression level of AcAT (acetylated alpha tubulin) protein in a biological sample isolated from a subject of interest. According to claim 9,
When the expression level of the AcAT protein measured in the biological sample is higher than that of the control group, it is predicted that anticancer drug resistance has occurred or is likely to occur in the target subject. According to claim 9,
The anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, cediranib, lesta urtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, bevacizumab, cisplatin, cetuximab, viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib , estramustine, gemtuzumab ozogamicin, ibritumomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine, Doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacytidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludabine, enocitabine, flu Tamid, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pirarubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, exmestane, in the group consisting of aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine and carmustine A method comprising at least one selected species. An agent that reduces the activity or expression level of AcAT (acetylated alpha tubulin) protein; containing as an active ingredient, a pharmaceutical composition for the treatment of anticancer drug resistance in gastric cancer patients. According to claim 12,
The agent for reducing the activity or expression of the protein is at least one selected from the group consisting of compounds, peptides, peptide mimetics, aptamers, antibodies, and natural products that specifically bind to the AcAT protein or a portion thereof, composition. delete According to claim 12,
The composition may include a histone deacetylase 6 (HDAC6) protein, an expression promoter of HDAC6 protein, or an activator of HDAC6 protein; Or alpha-tubulin N-acetyltransferase (Alpha-tubulin N-acetyltransferase; aTAT1) protein activity or expression inhibitor or at least one selected from the group consisting of an expression inhibitor of the gene encoding the aTAT1 protein further further comprising , composition. According to claim 12,
The anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, cediranib, lesta urtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, bevacizumab, cisplatin, cetuximab, viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib , estramustine, gemtuzumab ozogamicin, ibritumomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine, Doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacytidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludabine, enocitabine, flu Tamid, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pirarubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, exmestane, in the group consisting of aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine and carmustine A composition comprising at least one selected species. According to claim 12,
The anticancer agent is for the treatment of gastric cancer, the composition. An agent that reduces the activity or expression level of AcAT (acetylated alpha tubulin) protein; containing as an active ingredient, a pharmaceutical composition for enhancing sensitivity to anticancer drugs in gastric cancer patients. According to claim 18,
The composition may include a histone deacetylase 6 (HDAC6) protein, an expression promoter of HDAC6 protein, or an activator of HDAC6 protein; Or alpha-tubulin N-acetyltransferase (Alpha-tubulin N-acetyltransferase; aTAT1) protein activity or expression inhibitors or at least one selected from the group consisting of expression inhibitors of the gene encoding the aTAT1 protein, a composition further comprising . According to claim 18,
The anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, cediranib, lesta urtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, bevacizumab, cisplatin, cetuximab, viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib , estramustine, gemtuzumab ozogamicin, ibritumomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine, Doxifluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacytidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludabine, enocitabine, flu Tamid, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pirarubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melpharan, altretmin, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, exmestane, in the group consisting of aminoglutesimide, anagrelide, navelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine and carmustine A composition comprising at least one selected species. An agent that reduces the activity or expression level of AcAT (acetylated alpha tubulin) protein; containing as an active ingredient, a pharmaceutical composition for preventing or treating anticancer drug-resistant gastric cancer. processing a candidate substance for gastric cancer cells or gastric cancer tissue expressing AcAT (acetylated alpha tubulin) protein in vitro; and
A method for screening a drug for treating anticancer drug resistance in gastric cancer patients, comprising measuring the activity or expression level of AcAT protein in the gastric cancer cells or gastric cancer tissues after treatment with the candidate substance. The method of claim 22,
When the activity or expression level of the AcAT protein measured after treatment of the candidate material is reduced, further comprising the step of determining the candidate material as a drug for treating resistance to anticancer drugs in gastric cancer patients, the screening method.

Images