CN106011291A - Molecular marker for diagnosing and treating hypopharyngeal cancer - Google Patents
Molecular marker for diagnosing and treating hypopharyngeal cancer Download PDFInfo
- Publication number
- CN106011291A CN106011291A CN201610602028.1A CN201610602028A CN106011291A CN 106011291 A CN106011291 A CN 106011291A CN 201610602028 A CN201610602028 A CN 201610602028A CN 106011291 A CN106011291 A CN 106011291A
- Authority
- CN
- China
- Prior art keywords
- zacn
- gene
- hypopharyngeal cancer
- albumen
- expression
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/118—Prognosis of disease development
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
Abstract
The invention discloses a ZACN gene and an expression product thereof capable of serving as molecular markers for diagnosing and treating hypopharyngeal cancer. The content of the ZACN gene and the expression product thereof in a hypopharyngeal tissue of a subject can be detected to judge whether the subject suffers from hypopharyngeal cancer or not or diagnose whether the subject has a risk of suffering from hypopharyngeal cancer or not. Expression of the ZACN gene can be suppressed to suppress cell proliferation and suppress cell migration and invasion by researching the condition of proliferation, migration, invasion and the like of hypopharyngeal cancer cells cultured in vivo, and research results show that the ZACN gene and the expression product thereof are potential drug targets for treating hypopharyngeal cancer.
Description
Technical field
The present invention relates to diagnosing tumor, treat, predict prognosis field, more particularly it relates to detection
ZACN is abnormal is the diagnosing tumor of means, prediction method of prognosis；And suppression ZACN gene or protein
Tumor therapeutic agent.
Background technology
Hypopharyngeal cancer is also known as hypopharyngeal carcinoma.Abroad account for the 5% of head-neck malignant tumor, account for whole body malignant tumor
0.3%.Domestic account for the 1.4% of head-neck malignant tumor, account for the 0.2% of whole body malignant tumor.Although hypopharyngeal cancer
Sickness rate is relatively low, but owing to its position is hidden, clinical symptoms occurs later, and biological property is severe again,
67%～during 92% patient assessment already at III, IV phase, and range of tumor is often underestimated, and prognosis is poor.
Although the diagnosis and treatment means of hypopharyngeal cancer are updating height at present, but hypopharyngeal cancer particularly advanced cases with hypopharyngeal carcinoma survival rate
But without significantly improving, reason is that hypopharyngeal cancer early diagnosis is the most difficult, reaches an advanced stage and is difficult to control to cancerous cell
Transfer.The early diagnosis of cancer and the control of cancer cell metastasis are focuses clinically, the generation of research tumor
And metastatic rule, find its mark and potentially contribute to the generation of accurate evaluation cancer, development, be conducive to treatment
The formulation of measure, improves the prognosis of patient.
Summary of the invention
An object of the present invention is that providing a kind of is examined by detection ZACN gene or protein expression difference
The method of disconnected hypopharyngeal cancer.
The two of the purpose of the present invention are that providing a kind of comes pre-by detection ZACN gene or protein expression difference
The method surveying hypopharyngeal cancer prognosis.
The three of the purpose of the present invention are that providing a kind of is treated by suppression ZACN gene or ZACN albumen
The method of hypopharyngeal cancer.
A kind of method that the four of the purpose of the present invention are to provide medicine for screening treatment hypopharyngeal cancer.
The five of the purpose of the present invention are to provide a kind of medicine for treating hypopharyngeal cancer.
To achieve these goals, present invention employs following technical scheme:
The product that the invention provides detection ZACN gene or ZACN albumen is preparing hypopharyngeal cancer diagnostic tool
In purposes.
The product that present invention also offers detection ZACN gene or ZACN albumen is pre-at preparation prediction hypopharyngeal cancer
Purposes in rear instrument.
Further, the product of described detection ZACN gene or ZACN albumen include detect ZACN gene or
The product of the expression of ZACN albumen.Described product include can in conjunction with the nucleic acid of ZACN gene or
Can be in conjunction with the material (such as antibody) of ZACN albumen.Described nucleic acid can detect the expression of ZACN gene
Level；Described material can detect the expression of ZACN albumen.
The product of the detection ZACN gene of the present invention can play it based on the known method using nucleic acid molecules
Function: as PCR, as Southern hybridization, Northern hybridization, dot blot, fluorescence in situ hybridization (FISH),
DNA microarray, ASO method, high-flux sequence platform etc..Use this product can qualitatively, quantitatively,
Or semi-quantitatively implement to analyze.
The nucleic acid being included in the said goods can be obtained by chemosynthesis, or by preparing from biomaterial
Containing expectation nucleic acid gene, then use be designed for amplification expectation nucleic acid primer amplification it obtain.
Further, described PCR method is known method, such as, and ARMS (Amplification Refractory
Mutation System, amplification do not answer abruptly-changing system) method, RT-PCR (reverse transcriptase-PCR) method, nesting
PCR method etc..The nucleic acid of amplification can be by using dot blotting hybridization method, Surface Plasmon Resonance (SPR
Method), PCR-RFLP method, B by means of in situ RT PCR, PCR-SSO (sequence specific oligonucleotide) method,
PCR-SSP method, AMPFLP (amplifiable fragment length polymorphism) method, MVR-PCR method and
PCR-SSCP (single strand conformation polymorphism) method detects.
Nucleic acid recited above includes the primer expanding ZACN gene, and the primer that product includes can pass through
Being prepared by chemosynthesis, the method that be those skilled in the art will know that by use is come suitably with reference to Given information
Ground design, and prepared by chemosynthesis.
In specific embodiments of the present invention, described nucleic acid is the amplimer used in QPCR experiment, institute
State shown in sequence such as SEQ ID NO.1 (forward sequence) and SEQ ID NO.2 (reverse sequence) of primer.
Nucleic acid recited above may also include probe, and described probe can be prepared by chemosynthesis, by making
Appropriately design with reference to Given information by the method that those skilled in the art will know that, and prepared by chemosynthesis,
Or the gene containing expectation nucleotide sequence from biomaterial preparation can be passed through, and use is designed for the amplification phase
Hope nucleotide sequence primer amplification it prepare.
The product of the detection ZACN albumen of the present invention can play its function based on the known method using antibody:
For example, it is possible to include ELISA, radioimmunoassay, immunohistochemical method, western blot etc..
The product of the detection ZACN albumen of the present invention includes antibody or its sheet of specific binding ZACN albumen
Section.Antibody or its fragment of any structure, size, immunoglobulin class, origin etc. can be used, as long as
It combines target protein.Antibody or its fragment that the detection product of the present invention includes can be monoclonal
Or it is polyclonal.Antibody fragment refers to an antibody part (Partial Fragment) retaining antibody to the combination activity of antigen
Or the peptide containing an antibody part.Antibody fragment can include F (ab ')2, Fab ', Fab, scFv (scFv),
The Fv (dsFv) of disulphide bonding or its polymer, dimerization V district (double antibody) or containing CDR
Peptide.The product of the detection ZACN albumen of the present invention can include the amino of encoding antibody or Encoding Antibody Fragment
The nucleic acid of the separation of acid sequence, comprises the carrier of this nucleic acid, and carries the cell of this carrier.
Antibody can be by well known to a person skilled in the art that method obtains.Such as, preparation retains whole or portion
The polypeptide dividing target protein or the mammalian cell expression vector integrating their polynucleotide of coding are as anti-
Former.Use after antigen-immunized animal, from through the animal adaptive immune cell of immunity fused bone myeloma cells with
Obtain hybridoma.Then antibody is collected from Hybridoma culture.Finally can be used as antigen by use
ZACN albumen or its part antibody to obtaining are implemented antigenic specificity purification and are obtained for ZACN albumen
Monoclonal antibody.Polyclonal antibody can be prepared as follows: with antigen-immunized animal same as above, from warp
The animal crossing immunity collects blood sample, isolates serum from blood, then uses above-mentioned antigen real to serum
Execute antigenic specificity purification.Can be by the antibody obtained with ferment treatment or the sequence of the antibody obtained by use
Information obtains antibody fragment.
The combination of label and antibody or its fragment can be implemented by method as commonly known in the art.Such as,
Can fluorescent marker protein or peptide as follows: clean protein or peptide with phosphate buffer, add with DMSO,
Buffer agent, etc. preparation dyestuff, then mixed solution, then at room temperature place 10 minutes.It addition, labelling can
The labelling kit of commodity in use, such as biotin labeling reagent box, as biotin labeling reagent box-NH2,
Biotin labeling reagent box-SH (DojindoLaboratories)；Alkali phosphatase enzyme mark test kit such as alkalescence phosphorus
Acid enzyme labelling test kit-NH2, alkali phosphatase enzyme mark test kit-SH (Dojindo Laboratories)；Peroxide
Compound enzyme labelling test kit such as peroxidase labelling test kit-NH2, peroxidase labelling test kit
-NH2(Dojindo Laboratories)；Phycobniliprotein labelling kit such as phycobniliprotein labelling kit
-NH2, phycobniliprotein labelling kit-SH, B-phycoerythrin labelling kit-NH2, B-phycoerythrin
Labelling kit-SH, R-PE labelling kit-NH2, R-PE labelling kit
SH(DojindoLaboratories)；Fluorescent labeling reagent box such as fluorescein labelling kit-NH2, HiLyte
Fluor (TM) 555 labelling kit-NH2, HiLyte Fluor (TM) 647 labelling kit-NH2 (Dojindo
Laboratories)；And DyLight 547 and DyLight647 (Techno Chemical Corp.), Zenon (TM),
Alexa Fluor (TM) antibody labeling test kit, Qdot (TM) antibody labeling test kit (Invitrogen
And EZ-label Protein Labeling Kit (Funakoshi Corporation) Corporation).In order to correctly
Labelling, it is possible to use suitable instrument detects the antibody through labelling or its fragment.
Sample as the detection product according to the present invention, it is possible to use the tissue such as obtained from biopsy experimenter
Sample or fluid.Sample is not particularly limited, as long as it is suitable to the mensuration of the present invention；Such as, it can include
Tissue, blood, blood plasma, serum, lymph fluid, urine, serous cavity liquid, spinal fluid, synovial fluid, aqueous humor, tear
Liquid, saliva or its fraction or treated material.
In specific embodiments of the present invention, described sample is from the tissue of experimenter.
In the present invention, " prognosis " refer to that tumor patient is by the suppression such as surgical procedure or alleviate tumor growth
After process or result.In this manual, prognosis can be to be suppressed by surgical procedure or alleviate tumor growth
Life state when latter 1,2,3,4,5,6,7,8,9,10,15,20 years or more long.Prognosis can
With by checking that the gene of biomarker i.e. ZACN albumen or coding ZACN albumen is predicted.Prognosis is pre-
Survey can be performed such that according to biomarker with or without, or be raised and lowered, determine the prognosis of patient
It is good or bad, or determines the probability of good prognosis or poor prognosis.
In the present invention, " prognosis bona " refers to suppressed or alleviation tumor life for patient by surgical procedure etc.
After length, patient (such as 3,5,6,7,8,9,10,15,20 years or longer) over a long time not danger
Anxious situation.Or, good prognosis can mean to survive in the most long-time, without transfer, without recurrence or without again
Send out.Such as, prognosis bona can mean at least 3 years or survival in especially at least 5 years, preferably without transfer
Or recurrence.The most preferred state of prognosis bona is the long-term survival without disease.As used herein, " pre-
Rear good " can also include any such state, wherein it appeared that disease such as transfer, but pernicious low and
Do not severely impact survival ability.
In the present invention, " prognosis mala " refers to that patient is by the suppression such as surgical procedure or alleviation tumor growth
After short-term (such as 1,2,3,4,5 years or shorter) in occur fatal condition.Or, poor prognosis is
Refer to death in such short-term, shift, recur or send out again.Such as, poor prognosis can mean at least 3
Year or Preventive or death in especially at least 5 years.
Prediction prognosis refers to predict process or the result of status of patient, and being not meant to can be with the accuracy of 100%
The process of prediction status of patient or result.Prediction prognosis refers to whether the probability determining some process or result increases
Add, and be not meant to be compared by situation about not occurring with some process or result determine generation some process
Or the probability of result.For the present invention, the level liter of ZACN gene or ZACN albumen in the present invention
In high patient, compared with the patient not showing this feature, more likely observe particular procedure or result.
Further, the product of described detection ZACN gene or ZACN albumen can be detection ZACN gene
Or the reagent of ZACN albumen, can also be to comprise the test kit of described reagent, chip, reagent paper etc., it is possible to
To be the high-flux sequence platform using described reagent.
Present invention also offers a kind of instrument diagnosing hypopharyngeal cancer, described instrument can detect ZACN gene or
The expression of ZACN albumen.Described instrument includes can be in conjunction with the nucleic acid of ZACN gene or can tie
Close the material (such as antibody) of ZACN albumen.Described nucleic acid can detect the expression of ZACN gene；
Described material can detect the expression of ZACN albumen.
Further, the character of described nucleic acid and described material is with noted earlier.
Further, the instrument of described diagnosis hypopharyngeal cancer includes but not limited to chip, test kit, reagent paper or high pass
Measure sequence platform；High-flux sequence platform is the instrument of a kind of special diagnosis hypopharyngeal cancer, along with high-flux sequence
The development of technology, will become the structure of the gene expression profile of a people and work the most easily.By contrast disease
Patient and the gene expression profile of normal population, the exception easily analyzing which gene is relevant to disease.Therefore,
The exception purposes that fall within ZACN gene relevant to hypopharyngeal cancer of ZACN gene is known in high-flux sequence,
Same within protection scope of the present invention.
Present invention also offers a kind of instrument predicting hypopharyngeal cancer prognosis, described prediction hypopharyngeal cancer prognostic tool includes
Can be in conjunction with the nucleic acid of ZACN gene or can be in conjunction with the material (such as antibody) of ZACN albumen.Described
Nucleic acid can detect the mRNA level in-site of ZACN gene；Described material can detect the expression of ZACN albumen
Level.
Further, the character of described nucleic acid and described material is with noted earlier.
Further, the instrument of described prediction hypopharyngeal cancer prognosis include but not limited to chip, test kit, reagent paper or
High-flux sequence platform；High-flux sequence platform is the instrument of a kind of special diagnosis hypopharyngeal cancer, along with high flux
The development of sequencing technologies, will become the structure of the gene expression profile of a people and work the most easily.By right
Ratio Disease and the gene expression profile of normal population, the exception easily analyzing which gene is relevant to disease.
Therefore, in high-flux sequence, know that the exception of ZACN gene is relevant to hypopharyngeal cancer fall within ZACN gene
Purposes, equally within protection scope of the present invention.
The ammonia that the anti-ZACN antibody used in detection product, the diagnostic tool of the present invention or its fragment are identified
The number of base acid is not particularly limited, as long as antibody can be in conjunction with ZACN.When antibody is as curative
During thing, preferably it is capable of identify that aminoacid as much as possible, as long as it can suppress ZACN function.Anti-
The amino acid whose number of body or its fragment identification is at least one, more preferably at least three.The immune globulin of antibody
White classification is unrestricted, can be IgG, IgM, IgA, IgE, IgD or IgY.
Present invention also offers a kind of diagnose hypopharyngeal cancer or prediction hypopharyngeal cancer prognosis method, described method include as
Lower step:
(1) sample of experimenter is obtained；
(2) ZACN gene or the expression of albumen in detection Samples subjects；
(3) the ZACN gene recorded or the expression of albumen are associated with the whether ill of experimenter.
(4) compared with the control, the expression of ZACN gene or albumen raises, then this experimenter is diagnosed
For hypopharyngeal cancer, or this experimenter is confirmed as prognosis mala.
Present invention also offers the Therapeutic Method of a kind of hypopharyngeal cancer, described method include suppress ZACN gene or
ZACN albumen.
Further, described method includes the expression suppressing ZACN gene, or the expression of suppression ZACN albumen
Or the activity of suppression ZACN albumen.
Present invention also offers the screening technique of a kind of tumour medicine, can be by tumor cell be being added test
Measure certain period after medicine or after tumor model animal is used testing drug ZACN gene or
The expression of ZACN albumen measures tumour medicine and improves the effect of tumor prognosis.More specifically, when
The expression of ZACN gene or ZACN albumen is when adding or reduce after using testing drug or extensive
During multiple normal level, this medicine optional is as the medicine improving tumor prognosis.
Present invention also offers a kind of containing ZACN gene or the medicine of the inhibitor of ZACN albumen.
Present invention also offers the application in the medicine of preparation treatment hypopharyngeal cancer of the above-mentioned inhibitor.
The ZACN gene of the present invention or the inhibitor of ZACN albumen are unrestricted, as long as can suppress
ZACN or relate to the expression of material or the activity of ZACN upstream or downstream pathway, and treatment tumor is had
The medicine of effect.
Further, described inhibitor includes antisensenucleic acids, dsRNA, ribozyme, fit, ZACN associated proteins
Fragment or antibody or its fragment.
" antisensenucleic acids " refers to containing the nucleic acid with the sequence of the mRNA complementation of coding ZACN.Antisensenucleic acids
Can by DNA, RNA or the two form.Antisensenucleic acids need not and the mRNA100% of target ZACN
Complementary.Antisensenucleic acids can contain Non-complementary bases, as long as it can specific hybrid under strict conditions.
When antisensenucleic acids is introduced cell, it combine target polynucleotide and suppression is transcribed, RNA processing, translation or
Stability.In addition to antisense polynucleotides, antisensenucleic acids also includes polynucleotide analogies, and it contains through repairing
The main chain of decorations and 3 ' and 5 ' end portion.Such antisensenucleic acids can come just according to ZACN sequence information
When design use well known to a person skilled in the art that method generates.
" dsRNA " refers to, containing duplex-RNA constructs, carry out inhibition of gene expression by RNA interference (RNAi)
RNA, including siRNA (short interfering rna) and shRNA (short hairpin RNA).DsRNA need not
With the homology that target-gene sequence has 100%, as long as it can suppress expression of target gene.For stabilisation
Or other purpose, a part of DNA of dsRNA can be substituted.Preferably, siRNA is 21-23
The double-stranded RNA of individual base.SiRNA can be by well known to a person skilled in the art prepared by method, such as
By chemosynthesis or as the analog naturally occurring RNA.ShRNA is to have hair clip corner (hairpin
Turn) Short interfering RNA of structure.ShRNA can by well known to a person skilled in the art prepared by method,
Such as by chemosynthesis or by the DNA of coding shRNA is introduced cell expressible dna.
" ribozyme " refers to the RNA with catalysis activity, and it can cut, pastes, insert and transfer RNA.
The structure of ribozyme can include tup, hair clip etc..
" fit " refers to combine the nucleic acid of something such as protein.Fit can be RNA or DNA.Nucleic acid
Form can be double-strand or strand.Fit infinite in length system, if it can specific binding target molecule,
Can by such as 10 to 200 nucleotide, preferably 10 to 100 nucleotide, more preferably 15 to 80
Nucleotide, even more preferably 15 to 50 nucleotide compositions.Fit can use those skilled in the art
Known method selects.It is for instance possible to use SELEX (part carried out by exponential form enrichment be
System is evolved).
" the protein-bonded fragment of ZACN " refers to combine ZACN and the albumen of suppression ZACN enforcement original function
The fragment of matter.
The medicine of the present invention can be administered alone as medicine or use together with other medicines.Can be with the present invention
The other medicines used together of medicine unrestricted, as long as it does not damage the therapeutic of the present invention or preventative medicine
The effect of thing, it is preferred that the medicine for treatment or prophylaxis of tumours can include such as alkylating agent, all
As ifosfamide, cyclophosphamide, dacarbazine, temozolomide, nimustine, busulfan, procarbazine,
Melphalan and Ranimustine；Antimetabolite, such as enocitabine, capecitabine, carmofur, cladribine,
Gemcitabine, cytosine arabinoside, cytosine arabinoside octadecyl phosphate (cytarabine ocfosfate), ftorafur,
UFT, ftorafur gimeracil oteracil potassium, doxifluridine, hydroxyurea, fluorouracil,
Fludarabine, pemetrexed, pentostatin, mercaptopurine and methotrexate；Plant alkaloid, such as Yi Li
For health, etoposide, sobuzoxane, docetaxel, nogitecan, Pa Litasai, vinorelbine, Changchun
Pungent and the vinblastine in ground；Antitumor antibiotic, such as actinomycin D, aclarubicin, amrubicin, Yi Da ratio
Star, epirubicin, zinostatin stimalamer, daunorubicin, doxorubicin, pirarubicin, rich come mould
Element, peplomycin, ametycin and mitoxantrone；Medicine based on platinum, such as oxaliplatin, carboplatin,
Cisplatin and nedaplatin；Hormonal medicaments, such as Anastrozole, exemestane, estramustine, ethinylestradiol, chlorine
Ground progesterone, goserelin, tamoxifen, dexamethasone, toremifene, bicalutamide, flutamide, bold and vigorous Buddhist nun
Song Long, fostestrol, mitotane, methyltestosterone, medroxyprogesterone, mepitiostane, leuprorelin and letrozole；Raw
Thing reaction dressing agent, such as interferon-ALPHA, interferon beta, interferon gamma, interleukin, ubenimex, dry BCG,
And lentinan；And molecular targeted agents, such as imatinib (imatinib), gefitinib (gefitinib), Ji
Nurse monoclonal antibody, ozogamicin, Tamibarotene, trastuzumab, tretinoin, bortezomib (bortezomib),
With Rituximab etc..
The medicine of the present invention can be prepared as various dosage form as required.Include but not limited to, percutaneous, mucosa, nose,
Buccal, Sublingual or the tablet of per os use, solution, granule, patch, unguentum, capsule, aerosol
Or suppository.
The route of administration of the medicine of the present invention is unrestricted, as long as it can play desired therapeutic effect or pre-preventive effect
Fruit, includes but not limited to intravenous, intraperitoneal, ophthalmic, intra-arterial, in lung, is administered orally, in vesicle,
Intramuscular, tracheal strips, subcutaneous, by skin, by pleura, local, suck, by mucosa, skin
Skin, the intestines and stomach, intraarticular, in ventricle, rectum, vagina, in skull, in urethra, in liver, in tumor.At certain
In the case of Xie, can systematically be administered.It is to be administered partly in some cases.
The dosage of the medicine of the present invention is unrestricted, as long as obtaining desired therapeutic effect or preventive effect,
Appropriate determination can be carried out according to symptom, sex, age etc..The medicine of the present invention or the agent of prophylactic agent
Amount can use such as therapeutic effect or preventive effect to disease to determine as index.
In the context of the present invention, " diagnosis hypopharyngeal cancer " both includes judging that experimenter has suffered from hypopharynx
Cancer, also include judging whether experimenter exists the risk suffering from hypopharyngeal cancer.
" treatment " used herein contains treatment phase in the mammal such as mankind suffering from relevant disease or disease
The disease closed or morbid state, and include:
(1) prevention disease or morbid state occur in mammal, especially susceptible in institute when this mammal
State morbid state, but be not yet diagnosed when suffering from this morbid state；
(2) suppression disease or morbid state, i.e. stop it to occur；Or
(3) disease or morbid state are alleviated, even if disease or morbid state disappear.
Term " is treated " and is usually directed to treat the mankind or animal (such as, applied by veterinary), wherein can reach certain
Some intended therapeutic effect, such as, the development (including reducing development speed, making development stop) of suppression disease,
Improve disease and cure disease.Also include the treatment as preventive measure (such as prevention).To the most not developing into
Disease but have develops into the purposes of the dangerous patient of this disease, is also included within during term " treats ".
Advantages of the present invention and beneficial effect:
The present invention is found that a kind of molecular marker diagnosing hypopharyngeal cancer, use this molecular marker can under
The early stage that pharyngeal cancer occurs can be used as judging, it is provided that the survival rate of patient.
It addition, by the prognosis predicting patient, the present invention can provide significant information to determine to control for patient
Treat scheme policies.
The medicine of the inhibitor including ZACN gene or albumen of the present invention can be used as new hypopharyngeal cancer
Medicine.
Accompanying drawing explanation
Fig. 1 shows the differential expression utilizing QPCR to detect ZACN gene in mRNA level in-site；
Fig. 2 shows the differential expression utilizing immunoblotting to detect ZACN gene on protein level；
Fig. 3 show utilize QPCR detect the siRNA jamming effectiveness to ZACN gene expression；
Fig. 4 shows the jamming effectiveness utilizing immune-blotting method siRNA to ZACN gene expression；
Fig. 5 shows that utilizing MTT to detect suppresses the ZACN gene expression impact on hypopharyngeal cancer cell proliferation；
Fig. 6 shows the suppression ZACN protein function impact on hypopharyngeal cancer cell proliferation.
Specific embodiment
The present invention is further detailed explanation with embodiment below in conjunction with the accompanying drawings.Following example are only used for
The bright present invention rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in embodiment is logical
Often according to normal condition, such as Sambrook et al., molecular cloning: laboratory manual (New York:Cold
Spring HarborLaboratory Press, 1989) condition described in, or according to the bar proposed by manufacturer
Part.
Embodiment 1 gene chip screening difference expression gene
1, draw materials:
The primary Hypopharyngeal Cancer Patients excision cancerous tissue of the 8 routine dissections same period, separately takes 9 examples non-
The hypopharynx normal mucosa setup action comparison of hypopharyngeal cancer Disease.Under the equal verified by postoperative pathology of all cancerous tissues is
Pharyngeal cancer.Whole primary Hypopharyngeal Cancer Patients the most non-preoperative row Radiotherapy chemotherapy, all cases complete clinical data.
2, the acquisition of RNA is organized
Trizol one-step method is used to extract total tissue RNA.
3, RNA purity and the mensuration of concentration
Take RNA solution 1 μ l, Instrument measuring OD260, OD280, RNA concentration be OD260 value ×
Extension rate × 40/1000, calculates OD260/OD280, and it is high that ratio represents RNA solution purity at 1.7-2.0,
Few containing impurity such as protein ,-20 DEG C of preservations.
4, RNA integrity detection
(1) 2 μ l RNA sample row 1.5% agarose gel electrophoresis (80v, 15min) are taken；
(2) after separating zone, genefinder dyes, and observes electrophoresis zone under blue light；
(3) as 28s/18s about 2:1, illustrate that RNA is stable without degraded.
5, high flux transcript profile order-checking
5.1RNA-seq reads section location
First low-quality reading section is removed and obtain cleaning reading section, then utilize TopHat v1.3.1 by cleansing tablet
Section is mated with UCSC H.sapiens reference genome (hg19), H.sapiens UCSC hg19 version pre-
The index first built is downloaded from TopHat homepage, and as with reference to genome, utilizes TopHat and genome
Timing, it is allowed to each reading section (defaulting to 20) has multiple coupling site, most 2 mispairing.TopHat root
Possible shearing site storehouse is set up, according to these shearing site storehouses according to exon region and GT-AG shear signal
The reading section not navigating to genome is navigated on genome.The system of the TopHat method that we use is write from memory
Recognize parameter.
5.2 transcript abundance assessments
The reading segment file matched is by Cufflinks v1.0.3 process, and Cufflinks v1.0.3 is by RNA-seq sheet
Hop count mesh is standardized calculating the relative abundance of transcript.FPKM value refers in each million order-checking fragments
Match the segment number of the exon region of specific gene 1kb length.Calculated by Bayesian inference method
The confidence interval of FPKM estimated value.The GTF comment file of the reference that Cufflinks uses is from Ensembl number
(Homo_sapiens.GRCh37.63.gtf) is downloaded according to storehouse.
The detection of 5.3 difference expression genes
Ensembl GTF file and the original document mated by TopHat of download are transferred to Cuffdiff,
Cuffdiff uses original matching files to re-evaluate the gene expression abundance of the transcript listed in GTF file, inspection
Survey differential expression.Q value ＜ 0.01, test display is only had successfully more just to be considered in Cuffidff exports
It it is differential expression.
6, result
RNA-Sep result shows, filters out 211 differences between hypopharyngeal cancer tissue and normal control tissue altogether
Expressing gene, the gene 54 that wherein expression raises, the gene 157 that expression is lowered.
The difference expression gene filtered out verified by embodiment 2 large sample
Result based on the order-checking of the early stage high flux transcript profile degree of depth, according to the size of P value, we select
ZACN gene is verified.
1, sample collection
Collect hypopharyngeal cancer according to the method for embodiment 1 and organize 45 examples, normal control tissue 50 example.
2, verify in mRNA level in-site
2.1 extract tissue RNA
Step is with embodiment 1.
2.2 reverse transcription
Reverse transcription uses Primescript 1stStrand cDNA synthesis kit test kit, operating procedure is entered as follows
OK:
(1) in microcentrifugal tube, following reaction liquid is added, as shown in table 1:
Table 1 reaction liquid
| Reagent | Dosage |
| RNA | 2.0μg |
| dNTP | 1.0μl |
| Oligo(dT) | 2.0μl |
| Rnase free dH2O | Add to 10.0 μ l |
Hatch 5min, be rapidly cooled to 4 DEG C for (2) 70 DEG C；
In microcentrifugal tube, add following reaction reagent, make reaction system:
| Reagent | Dosage |
| 5x1st Strand Synthesis Buffer | 4.0μl |
| PrimeScript RTase | 1.0μl |
| RNase Inhibitor | 1.0μl |
| Rnase free dH2O | 4.0μl |
Shaking gently, after being quickly centrifuged, 42 DEG C of reaction 1h, 70 DEG C of 10min terminate reacting, 4 DEG C of coolings ,-20 DEG C
Preserve.
Use SYBP Premix Ex TapTMII test kit, at Eppendorf Real-time pcr analysis instrument
Carrying out, concrete operations are as follows:
(1) following PCR reactant liquor is prepared on ice:
| Reagent | Dosage |
| SYBR | 10.0μl |
| Forward primer | 1.0μl |
| Reverse primer | 1.0μl |
| cDNA | 2.0μl |
| ddH2O | 6.0μl |
| Total amount | 20.0μl |
Primer sequence design is as follows:
ZACN gene:
5’-GCGATGGAGTTAGAGTTC-3’(SEQ ID NO.1)；
5’-TGGGTCTTCAGATCATAAAC-3’(SEQ ID NO.2)
β-actin:
5’-GTGGGGCGCCCCAGGCACCA-3’(SEQ ID NO.3)；
5’-CTCCTTAATGTCACGCACGATTT-3’(SEQ ID NO.4)
(2) upper machine, performs following program: 95 DEG C of denaturations 3min；95 DEG C of degeneration 15s.59 DEG C of annealing 20s,
72 DEG C extend 20s, totally 40 circulations.
Result relative quantification method, using formula 2-△ΔctCalculate.Experiment is repeated 3 times.
△ ct=ct (A)-ct (β-actin)
△ Δ ct=△ ct (experimental group)-△ ct (matched group)
Result such as Fig. 1 shows, compared with normal control tissue, and the mRNA of ZACN gene in hypopharyngeal cancer tissue
Level substantially raises, and difference has statistical significance (P < 0.05).
3, verify on protein level
Extract each histone according to RIPA protein cleavage liquid kit description, use BCA protein concentration to survey
Determine protein concentration in test kit detection sample.With conventional Western-blot method detection ZACN albumen change,
Each group experiment is all repeated 3 times, and with β-actin as internal reference, does ZACN protein band absorbance quantitative analysis,
Expression represents with the ratio of ZACN albumen/β-actin absorbance.
Result such as Fig. 2 shows, compared with normal control tissue, in hypopharyngeal cancer tissue, ZACN protein level shows
Writing and increase, difference has statistical significance (P < 0.05).
Embodiment 3 suppresses ZACN gene expression
1, siRNA design synthesis
SiRNA sequence for ZACN:
SiRNA-ZACN:
Positive-sense strand is 5 '-AUCAUAAACUACGUAUUCCCU-3 ' (SEQ ID NO.5)
Antisense strand is 5 '-GGAAUACGUAGUUUAUGAUCU-3 ' (SEQ ID NO.6)；
Above siRNA sequence and negative control siRNA sequence (siRNA-NC) are by Shanghai Ji agate pharmacy skill
Art company limited provides.
2, the cultivation of hypopharyngeal cancer cell and transfection
2.1 cells are cultivated
Hypopharyngeal cancer FADU cell uses containing 10% hyclone (FBS), penicillin 100U/ml, chain
RPMI 1640 culture medium of mycin 100 μ g/ml is placed in 37 DEG C, 5%CO2, train under saturated humidity environment
Support.
2.2 cell transfecting
(1) first 24 hours are transfected, at 500 μ l nonreactive inoculation of medium 0.5-2*105Individual cell, during transfection
Cell degrees of fusion is 30-50%.Cell dissociation is mixed completely during bed board, it is to avoid cell accumulation grows.
(2) siRNA (the final concentration of 33nM of transfectional cell, pressure-vaccum gently are diluted with 50 μ l Opti-MEM
3-5 mixing.
(3) overturn mixing transfection reagent gently, dilute 1 μ l Lipofectamine2000 with 50 μ l Opti-MEM
3-5 mixing of pressure-vaccum, left at room temperature 5min gently.
(4) mixing transfection reagent and siRNA diluent, gently 3-5 mixing of pressure-vaccum, left at room temperature 20min.
(5) during transfection composite joins 24 porocyte plates, 100 μ l/ hole, front and back jog cell plates mix homogeneously.
(6) cell plates are placed in 37 DEG C, 5%CO2Incubator is cultivated 18-48 hour.After transfecting 4-6 hour
Interchangeable fresh culture medium.
3, the jamming effectiveness of QPCR experiment detection siRNA is utilized
3.1 extract cell total rna utilizes conventional method to operate.
3.2 reverse transcription
Step is with embodiment 2.
3.3QPCR
Step is with embodiment 2.
3.4 result
Result is as it is shown on figure 3, compared with siRNA-NC group, siRNA-ZACN can effectively suppress ZACN
The expression of gene mRNA, difference has statistical significance (P < 0.05).
4, the jamming effectiveness of Western blot experiment detection siRNA is utilized
Step is with embodiment 2.
As shown in Figure 4, compared with siRNA-NC group, siRNA-ZACN can effectively suppress ZACN to result
The expression of albumen, difference has statistical significance (P < 0.05).
The expression of the embodiment 4ZACN gene mensuration to hypopharyngeal cancer ability of cell proliferation
1, step
Carry out cell transfecting according to the step of embodiment 2, by the transfection cell dissociation of 48 hours, be inoculated in 96
In well culture plate, 5000, every hole cell, every hole 200 μ L cell suspension, cultivate 24h, 48,72 respectively
After, with the person that is not added with cell for blank group, every hole adds 20 μ L MTT (0.5mg/mL), incubates in incubator
Carefully sucking liquid in hole after educating 4h, every hole adds 100 μ L DMSO, vibration 10～20min, makes crystallization
Thing fully dissolves.Enzyme-linked immunosorbent assay instrument measures each hole OD570 value, is respectively horizontal stroke with time and absorbance
Vertical coordinate, draws cell growth curve.Every porocyte arranges 3 multiple holes.This experiment is repeated 3 times.
2, result
Result is as it is shown in figure 5, compared with transfection siRNA-NC group, transfection siRNA-ZACN group cell increases
Growing slowly, difference has statistical significance (P < 0.05).Above-mentioned test result indicate that, ZACN gene expression
Promote the propagation of hypopharyngeal cancer cell.
In embodiment 5 hypopharyngeal cancer cell antibody and experiment
1, step:
Hypopharyngeal cancer cell FADU is inoculated in 96 porocyte culture plates, every hole 2*103Individual cells/well/200 μ l,
It is handled as follows after cell attachment:
Experimental group 1 (matched group): add unrelated monoclonal antibody (1:50) in hypopharyngeal cancer cell；
Experimental group 2: add anti-human ZACN monoclonal antibody (1:50) in hypopharyngeal cancer cell.
By cell at 37 DEG C, 5%CO2After incubator hatches 24 hours, according to Brd U cell proliferation reagent
The description of box (Chemicon International), measures cell proliferation rate.
2, statistical method
Experiment is all according to being repeated 3 times, and result data is all to come in the way of mean+SD
Representing, using SPSS13.0 statistical software to carry out statistical analysis, difference between the two uses t inspection,
Think when P < has statistical significance when 0.05.
3, result
Result as shown in Figure 6, compared to matched group, adds the group cells proliferation slowed down of anti-human ZACN monoclonal antibody.
Above-mentioned test result indicate that, the function of suppression ZACN albumen can suppress hypopharyngeal cancer cell proliferation.
Embodiment 6 detects the impact of ZACN gene expression cell migration
1, experimental procedure is migrated
(1) by the cell dissociation after transfection 24h, adjusting cell density is 105/ml；
(2) the 24 every holes of porocyte culture plate add 600 μ L import hyclone (FBS), and Transwell is little
24 porocyte culture plate apertures are inserted in room, little indoor each 100 μ L RPMI 1640 culture medium of addition of Transwell
Cell suspension；
(3) 24 porocyte culture plates hatch 24h, are taken out from 24 porocyte culture plates by Transwell cell,
Discarding culture medium, PBS rinses Transwell cell film portion attached cell；
(4) taking new 24 porocyte culture plates, ((1:1) 600 μ L, will to add methanol/PBS mixed liquor in aperture
Transwell cell puts into aperture, makes methanol/PBS mixed liquor immerse from lower room (24 porocyte culture plate aperture)
Upper room, room temperature stands 15min；
(5) methanol/PBS mixed liquor in room and 24 porocyte culture plate apertures is discarded on Transwell cell,
Lower room adds 600 μ L methanol, and Transwell cell is inserted aperture, makes methanol from (the 24 porocyte trainings of lower room
Support plate aperture) room in immersions, room temperature standing 15min；
(6) discard methanol, under room temperature, be dried 15-30min；
(7) take 0.1% crystal violet dye liquor 600 μ L and instill 24 porocyte culture plate apertures, Transwell is little
Aperture dyeing 15min is inserted in room；
(8) discard 0.1% crystal violet dye liquor, distilled water flushing Transwell cell 15min, dry, micro-
Microscopic observation, takes the different visual field and takes pictures counting, and experiment is repeated 3 times.
2, result
Under transfection siRNA-NC group and the transfection each visual field of siRNA-ZACN group, average mobility cell number divides
Not Wei (167.45 ± 14.98) individual and (79.86 soil 8.34) individual, difference has statistical significance (P < 0.05).
Above-mentioned test result indicate that, ZACN gene expression promotes the migration of hypopharyngeal cancer cell.
Embodiment 7 detects the ZACN gene expression impact on cell invasion
1, Matrigel step
Matrigel is taken out from-20 DEG C of refrigerators before experimentTMMatrigel, ice chest melts, takes MatrigelTMSubstrate
Glue is mixed in 1:6 ratio with RPMI 1640 culture medium, makes mixed liquor and adds room on Transwell cell,
Every hole 45 μ L, inserts Transwell cell 24 porocyte culture plates, is transferred to 37 DEG C of 5%CO2Cultivate
Case hatches 30min, subsequent cell inoculation and cultivation operation with above-mentioned Cell migration assay.After cultivating 24h,
Discard Pei Ji, wipe indoor Matrigel on Transwell cell gently away with cotton swabTMMatrigel, does not damage little
Room counterdie, the same cell transfer experiments of remaining operation.This experiment is repeated 3 times.
2, result
Under transfection siRNA-NC group and the transfection each visual field of siRNA-ZACN group, average invasion and attack cell number divides
Not Wei (54.14 ± 6.46) individual and (26.25 soil 3.69) individual, difference has statistical significance (P < 0.05).
Above-mentioned test result indicate that, ZACN gene expression promotes the invasion and attack of hypopharyngeal cancer cell.
The detection of embodiment 8 body outer clone Forming ability
1, step
(1) making cell suspension after the cell dissociation after transfection 24h, cell counting count board counts；
(2) the six every holes of porocyte culture plate add 2ml 10%FBS-RPMI 1640 culture medium, by 500
/ porocyte density inoculating cell suspension, mixes gently；
(3) six porocyte culture plates move to 37 DEG C of 5%CO2Incubator is hatched 10 days, within every 3 days, changes and cultivates
Base 1 time, each 2ml/ hole；
(4) after cultivation terminates, discarding culture medium, PBS carefully cleans 3 times, each 5min, room
Temperature is dried, and methanol fixes 15min, discards methanol, and air at room temperature is dried 20min；
(5) taking 0.1% crystal violet dye liquor and add Tissue Culture Plate, every hole adds 1ml, and dye 15min；
(6) reclaiming 0.1% crystal violet dye liquor, Tissue Culture Plate distilled water cleans 15min；
(7) observation of taking pictures counts, and repeats to test 3 times.
2, result
It is individual that transfection siRNA-NC group average colony forms number respectively (175.30 ± 16.45), transfection
It is that (78.00 soil 5.56) is individual that siRNA-ZACN group cell average colony forms number.Above-mentioned experimental result table
Bright, ZACN gene expression promotes the one-tenth tumor ability of hypopharyngeal cancer cell.
The explanation of above-described embodiment is only intended to understand the method for the present invention and core concept thereof.It is right to it should be pointed out that,
For those of ordinary skill in the art, under the premise without departing from the principles of the invention, it is also possible to the present invention
Carrying out some improvement and modification, these improve and modify also by the protection domain falling into the claims in the present invention.
Claims (10)
1. the product of detection ZACN gene or ZACN albumen is pre-at preparation diagnosis hypopharyngeal cancer or prediction hypopharyngeal cancer
After instrument in application.
Application the most according to claim 1, it is characterised in that described detection ZACN gene or ZACN
The product of albumen includes the product detecting the expression of ZACN gene or ZACN albumen.
Application the most according to claim 1 and 2, it is characterised in that described product includes can be in conjunction with
The nucleic acid of ZACN gene or can be in conjunction with the material of ZACN albumen；Described nucleic acid can detect ZACN base
The expression of cause；Described material can detect the expression of ZACN albumen.
Application the most according to claim 3, it is characterised in that described nucleic acid is to make in real-time quantitative PCR
The primer of specific amplified ZACN gene as shown in SEQ ID NO.1 and SEQ ID NO.2.
5. the instrument diagnosing hypopharyngeal cancer or prediction hypopharyngeal cancer prognosis, it is characterised in that described instrument includes
The instrument of the expression of ZACN gene or ZACN albumen can be detected.
Instrument the most according to claim 5, it is characterised in that described instrument includes can be in conjunction with ZACN
The nucleic acid of gene or can be in conjunction with the material of ZACN albumen；Described nucleic acid can detect the table of ZACN gene
Reach level；Described material can detect the expression of ZACN albumen.
Instrument the most according to claim 6, it is characterised in that described nucleic acid is to make in real-time quantitative PCR
The primer of specific amplified ZACN gene as shown in SEQ ID NO.1 and SEQ ID NO.2.
8. the medicine treating hypopharyngeal cancer, it is characterised in that described pharmaceutical pack contains ZACN gene or ZACN
The inhibitor of albumen.
Medicine the most according to claim 8, it is characterised in that described inhibitor can suppress ZACN or
Relate to expression or the activity of the material of ZACN upstream or downstream pathway.
10. the application in the medicine of preparation treatment hypopharyngeal cancer of the inhibitor described in claim 8 or 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610602028.1A CN106011291B (en) | 2016-07-27 | 2016-07-27 | A kind of molecular marker of diagnosis and treatment hypopharyngeal cancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610602028.1A CN106011291B (en) | 2016-07-27 | 2016-07-27 | A kind of molecular marker of diagnosis and treatment hypopharyngeal cancer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106011291A true CN106011291A (en) | 2016-10-12 |
| CN106011291B CN106011291B (en) | 2019-09-10 |
Family
ID=57114274
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610602028.1A Active CN106011291B (en) | 2016-07-27 | 2016-07-27 | A kind of molecular marker of diagnosis and treatment hypopharyngeal cancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106011291B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110951880A (en) * | 2019-12-30 | 2020-04-03 | 西安交通大学医学院第二附属医院 | Application of reagent for detecting lncRNA marker of hypopharynx cancer in preparation of product for diagnosing hypopharynx cancer |
-
2016
- 2016-07-27 CN CN201610602028.1A patent/CN106011291B/en active Active
Non-Patent Citations (4)
| Title |
|---|
| BASYUK E等: "The candidate tumor suppressor gene ZAC is involved in keratinocyte differentiation and its expression is lost in basal cell carcinomas", 《MOL CANCER RES》 * |
| HU LI等: "Aberrantly expressed genes and miRNAs in human hypopharyngeal squamous cell carcinoma based on RNA sequencing analysis", 《ONCOLOGY REPORTS》 * |
| KOY S等: "Loss of expression of ZAC/LOT1 in squamous cell carcinonas of head and neck", 《HEAD NECK》 * |
| 李科宇等: "非小细胞肺癌ZAC基因的表达及临床意义", 《临床肺科杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110951880A (en) * | 2019-12-30 | 2020-04-03 | 西安交通大学医学院第二附属医院 | Application of reagent for detecting lncRNA marker of hypopharynx cancer in preparation of product for diagnosing hypopharynx cancer |
| CN110951880B (en) * | 2019-12-30 | 2022-08-09 | 西安交通大学医学院第二附属医院 | Application of reagent for detecting lncRNA marker of hypopharynx cancer in preparation of product for diagnosing hypopharynx cancer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106011291B (en) | 2019-09-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105969901B (en) | Purposes of the MS4A6A as Huppert's disease diagnosis and treatment marker | |
| CN105886659A (en) | DSTN gene and expression product thereof as diagnosis and treatment target of endometrial cancer | |
| CN106222259B (en) | A kind of molecular marker of diagnosis and treatment Huppert's disease | |
| CN106906290A (en) | CDSN as Dendritic cell diagnosis and treatment target | |
| CN106244705B (en) | Application of the ERC1 in preparation diagnosis or treatment hypopharyngeal cancer tool | |
| CN106011288B (en) | Huppert's disease diagnosis and treatment marker and its application | |
| CN106191283B (en) | The diagnosis and treatment product of Huppert's disease biomarker | |
| CN105907879A (en) | Endometrial cancer biological marker | |
| CN106011260B (en) | A kind of molecular marker of diagnosis and treatment carcinoma of endometrium | |
| CN106011291A (en) | Molecular marker for diagnosing and treating hypopharyngeal cancer | |
| CN105886660A (en) | Application of UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) as endometrial cancer diagnosis and treatment marker | |
| CN105969904B (en) | Huppert's disease biomarker | |
| CN106011289A (en) | Application of LILRA2 gene and expression product thereof as diagnosis and treatment target of multiple myeloma | |
| CN106947818A (en) | A kind of molecular marker of diagnosis and treatment adenocarcinoma of colon | |
| CN106947820A (en) | Purposes of the VCAN in adenocarcinoma of colon diagnosis and treatment | |
| CN106148531A (en) | Hypopharyngeal cancer biomarker | |
| CN106381329B (en) | The diagnosis and treatment target of C18orf8 gene and its expression product as hypopharyngeal cancer | |
| CN106119358B (en) | The diagnosis and treatment product of carcinoma of endometrium biomarker | |
| CN106119399B (en) | Purposes of the ZNF385C as hypopharyngeal cancer diagnosis and treatment marker | |
| CN107034270B (en) | CLIC3 as diagnosis and treatment target of lung adenocarcinoma | |
| CN106011286A (en) | Hypopharyngeal carcinoma diagnosis and treatment marker and application thereof | |
| CN106119400A (en) | C2CD4C application in preparation diagnosis and treatment hypopharyngeal cancer product | |
| CN105969903B (en) | The diagnosis and treatment product of hypopharyngeal cancer biomarker | |
| CN107034272A (en) | Applications of the CXCL2 in diagnosis or treatment adenocarcinoma of lung instrument is prepared | |
| CN106947819A (en) | Adenocarcinoma of colon diagnosis and treatment mark |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: Room 1210, Building 3, Ronghua Xintai Building, 10 Ronghua South Road, Yizhuang Economic and Technological Development Zone, Daxing District, Beijing Applicant after: Beijing Yang Shen biology information technology company limited Address before: 100080 Beijing city Haidian District Shanyuan Street No. 1 cubic court building room 3103 Applicant before: Beijing Yang Shen biology information technology company limited |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20211118 Address after: 266000 room 2503, Qianshan building, D2, Qingdao International Innovation Park Phase II, No.1 Keyuan Weiyi Road, Laoshan District, Qingdao City, Shandong Province Patentee after: Qingdao Yangshen biomedical Co.,Ltd. Address before: Room 1210, building 3, Ronghua Xintai building, 10 ronghua South Road, Yizhuang Economic and Technological Development Zone, Daxing District, Beijing Patentee before: BEIJING MEDINTELL BIOMED Co.,Ltd. |
|
| TR01 | Transfer of patent right |