KR101674874B1 - Icam-3 as biomarker for the prediction of resistance against anticancer agents and use thereof - Google Patents

Abstract

The present invention relates to a biomarker for predicting anticancer agents resistance and uses thereof, and more particularly, to a method for predicting anticancer drug resistance using ICAM-3 (intercellular adhesion molecule-3) A biomarker used and a diagnostic kit. The method, biomarker and kit of the present invention can promptly predict the occurrence of resistance to an anticancer drug in a patient who has received the anticancer drug treatment, so that it is possible to provide accurate basic information on planning and performing an effective protocol for future treatment by the clinician have.

Description

ICAM-3 as a Biomarker for Anticancer Drug Resistance Prediction and its Use {ICAM-3 AS BIOMARKER FOR THE PREDICTION OF RESISTANCE AGAINST ANTICANCER AGENTS AND USE THEREOF}

The present invention relates to a biomarker for predicting anticancer agents resistance and uses thereof, and more particularly, to a method for predicting anticancer drug resistance using ICAM-3 (intercellular adhesion molecule-3) A biomarker used and a diagnostic kit.

Cancer is one of the leading causes of death worldwide, and there are three main methods of treatment: surgical surgery, chemotherapy prescription, and radiation therapy. However, among these major treatment modalities, cancer and cancer treatment are often resistant to radiation and chemotherapeutic agents, leading to recurrence and metastasis, leading to death in cancer patients.

There are various theories about the resistance formation of cancer, but Meads et al . Advocated the environment-mediated drug resistance theory (Meads et al . , Nat. Rev. Cancer 9 (2009) 665-74). According to this theory, cancer cells that are sensitive to treatment die through apoptosis, while surviving through a new resistance mechanism due to microenvironment surrounding cancer cells, acquire inherent resistance (FIG. 1).

Therefore, in order to effectively treat cancer, in-depth research on resistance to radiation and chemotherapeutic treatment has become an important issue for cancer conquest.

KR 10-2013-7003284 A

A problem to be solved by the present invention is to provide an effective method for promptly predicting and determining whether a patient who has received an anticancer drug treatment can acquire tolerance to an anticancer drug and provide a biomarker and a kit for anticancer drug resistance prediction used in the method I would like to.

In order to solve the above-mentioned problems, the present invention provides a method for measuring the expression level of ICAM-3 in cancer cells isolated from cancer patients treated with an anticancer agent, measuring the expression level of ICAM-3 measured from cancer cells isolated from cancer patients before treatment with anti- A method of providing information for anticancer drug resistance prediction that the possibility of resistance to the anticancer drug is likely to occur when the expression level of ICAM-3 measured from the cancer cell isolated from the cancer patient after the treatment with the anticancer drug is increased compared with the current dose to provide.

The anticancer agent is preferably paclitaxel or vincristine.

The cancer cells are preferably lung cancer cells or cervical cancer cells.

The expression level of ICAM-3 is preferably measured by measuring the amount of mRNA or protein.

The amount of the mRNA may be measured by RT-PCR, northern blotting or a nucleotide microarray.

The amount of the protein is preferably measured by an antigen-antibody reaction.

The antigen-antibody reaction may be measured by Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry or protein microarray or aptamer, It is preferable that the detection is performed by using

The present invention also provides a biomarker for anticancer drug resistance prediction comprising ICAM-3.

The ICAM-3 is preferably a nucleic acid having the nucleotide sequence shown in SEQ ID NO: 1.

The ICAM-3 is preferably a protein having an amino acid sequence represented by SEQ ID NO: 2.

The present invention also provides a kit for anticancer drug resistance prediction comprising means capable of measuring the expression level of ICAM-3.

The means is preferably selected from the group consisting of a primer, a probe, an antibody, an antibody fragment, a microarray and an aptamer.

The method, biomarker and kit of the present invention can promptly predict the occurrence of resistance to an anticancer drug in a patient who has received the anticancer drug treatment, so that it is possible to provide accurate basic information on planning and performing an effective protocol for future treatment by the clinician have.

Fig. 1 schematically shows occurrence of tolerance to chemotherapy.
Fig. 2 shows the chemical structure of paclitaxel and vincristine, which are anti-cancer agents used in the examples of the present invention.
3 is a view for explaining the anticancer drug resistance-imparting effect of ICAM-3 inhibiting apoptosis.
FIG. 4 is a graph showing that the anticancer drug resistance-imparting effect of ICAM-3 is due to activation of Akt and ERK.
FIG. 5 is a graph showing that activation of CREB2 affects the anticancer drug resistance-enhancing effect of ICAM-3. FIG.
6 is a diagram illustrating the anticancer drug resistance mechanism by activation of ICAM-3-Akt / ERK-CREB2 in H460 cancer cells and radiation tolerance.
Fig. 7 is a graph showing the effect of ICAM-3 anticancer drug resistance-imparting effect in a mouse tumor graft model. Fig.
Fig. 8 schematically shows various functions of cancer cells of ICAM-8.

Hereinafter, the present invention will be described in detail.

ICAM-3 has been analyzed as a gene that confers radiation tolerance in the conventional studies involving the inventors of the present invention, and this gene has also been identified as having the function of increasing invasiveness and metastasis of cancer cells by the inventors of the present invention . Therefore, the inventors of the present invention have estimated that the ICAM-3 gene having such a function has been identified as a candidate gene capable of providing an important clue for induction of radiation therapy. In the present invention, the ICAM-3 gene has resistance against an anti- And the present invention has been completed.

Accordingly, the present invention provides a method for detecting the expression level of ICAM-3 in cancer cells isolated from cancer patients treated with an anticancer agent and comparing the amount of ICAM-3 detected from cancer cells isolated from cancer patients before the treatment with an anticancer agent, The present invention provides an information providing method for predicting anticancer drug resistance, which is judged to have a possibility of resistance to an anticancer agent when the expression level of ICAM-3 measured from cancer cells isolated from a cancer patient is increased.

In a preferred embodiment, the method of providing information for anticancer drug resistance prediction of the present invention may include the following steps:

A first step of preparing a sample from cancer cell tissue isolated from a cancer patient and measuring the amount of ICAM-3 expression from the sample;

A second step of preparing a sample from cancer cell tissue isolated from a cancer patient to which an anticancer drug is administered and measuring the amount of ICAM-3 expression from the sample;

Comparing the result of the first step with the result of the second step;

Providing information for determining that the possibility of resistance to an anticancer drug is high when the amount of expression of ICAM-3 in the second step is higher than the amount of expression of ICAM-3 in the first step through the comparison.

The anticancer agent is preferably paclitaxel or vincristine as shown in Fig.

The cancer cells are preferably lung cancer cells or cervical cancer cells.

The expression level of ICAM-3 is preferably measured by measuring the amount of mRNA or protein.

The amount of the mRNA may be measured by RT-PCR, northern blotting or a nucleotide microarray.

The amount of the protein is preferably measured by an antigen-antibody reaction.

The measurement by the antigen-antibody reaction can be performed by Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry or protein microarray, or detection by aptamer .

The present invention also provides a biomarker for anticancer drug resistance prediction comprising ICAM-3.

The biomarker of the present invention means a molecular marker for the purpose of predicting anticancer drug resistance. ICAM-3 used as the biomarker may be a nucleic acid having the nucleotide sequence shown in SEQ ID NO: 1 or a protein having the amino acid sequence shown in SEQ ID NO: 2. Also, it may be any base sequence or amino acid sequence having a homology of 70%, preferably 80%, more preferably 90% or more, to the base sequence of SEQ ID NO: 1 or the amino acid sequence of SEQ ID NO: 2.

The present invention also provides a kit for anticancer drug resistance prediction comprising means capable of measuring the expression level of ICAM-3.

The "ICAM-3" has the same meaning as the biomarker of the present invention described above.

The expression "expression amount of ICAM-3" means the amount of mRNA expression or protein expression of ICAM-3.

The means may be a known molecular biological means or immunochemical means.

In a preferred embodiment, the means may be a measuring means selected from the group consisting of primers, probes, antibodies, antibody fragments, microarrays and aptamers. Here, the primer, the probe, or the nucleic acid microarray corresponds to a means for measuring the expression amount of mRNA, and the antibody, the antibody fragment, the protein microarray or the aptamer corresponds to the means for measuring the expression amount of the protein.

The term "primer" refers to a single-stranded oligonucleotide capable of acting as a starting point for template-directed DNA synthesis under suitable conditions (ie, four different nucleoside triphosphates and polymerization enzymes) within a suitable buffer and a suitable temperature it means. The appropriate length of the primer may vary depending on various factors, such as temperature and use of the primer. In addition, the sequence of the primer does not need to have a sequence completely complementary to a partial sequence of the template, and it is sufficient that the primer has sufficient complementarity within a range capable of hybridizing with the template and acting as a primer. Therefore, the primer in the present invention does not need to have a perfectly complementary sequence to the nucleotide sequence of the ICAM-3 gene as a template, and it is sufficient if the primer has sufficient complementarity within the range capable of hybridizing with the gene sequence and acting as a primer .

Refers to a linear or linear oligomer of natural or modified monomer or linkages and can specifically hybridize to the nucleotide sequence of ICAM-3, which contains deoxyribonucleotides and ribonucleotides and is the target nucleotide, Or artificially synthesized. The probes according to the present invention may be single-stranded, preferably oligodeoxyribonucleotides. Probes of the invention can include natural dNMPs (i.e., dAMP, dGMP, dCMP and dTMP), nucleotide analogs or derivatives. In addition, the probe of the present invention may also include a ribonucleotide. For example, the probes of the present invention can be used in combination with a framework-modified nucleotide such as a peptide nucleic acid (PNA) (M. Egholm et al., Nature, 365: 566-568 (1993)), phosphorothioate DNA, phosphorodithioate DNA, phosphoamidate DNA, amide-linked DNA, MMI-linked DNA, 2'-O-methyl RNA, alpha-DNA and methylphosphonate DNA, sugar modified nucleotides such as 2'- 2'-O-alkyl DNA, 2'-O-allyl DNA, 2'-O-alkynyl DNA, hexose DNA, pyranosyl RNA, and anhydrohexy Tolyl DNA, and nucleotides with base modifications such as C-5 substituted pyrimidines wherein the substituents are fluoro-, bromo-, chloro-, iodo-, methyl-, ethyl-, vinyl-, formyl-, 7-deazapurines having a C-7 substituent (the substituents being fluoro, bromo, bromo, bromo, and iodo) Chloro-, Iodo-, Me For example, methyl, ethyl, vinyl, formyl, alkynyl, alkenyl, thioglyl, imidazolyl, pyridyl and the like), inosine and diaminopurine.

The "antibody" may include a polyclonal antibody, a monoclonal antibody, a recombinant antibody, or the like capable of specifically binding to the ICAM-3 protein.

The antibody is commercially available and can be used. Since the amino acid sequence of the ICAM-3 protein is known as described above, the antibody can be directly prepared using the amino acid sequence.

For example, in the case of a polyclonal antibody, an ICAM-3 antigen is injected into an animal and blood is collected from an animal to produce an antibody Such polyclonal antibodies can be produced from any animal species host, such as goat, rabbit, sheep, monkey, horse, pig, cow, dog, and the like . Monoclonal antibodies can be prepared using the hybridoma method (Kohler et al., European Jounal of Immunology, 6, 511-519, 1976) well known in the art or can be prepared using the phage antibody library Clackson et al., Nature, 352, 624-628, 1991, Marks et al., J. Mol. Biol., 222: 58, 1-597, 1991).

The term "antibody fragment" means a functional fragment of an antibody molecule. The functional fragment of an antibody molecule is an antibody fragment having at least an antigen binding function, for example, Fab, F (ab '), F ') 2 and Fv.

The "microarray" may be, for example, a nucleic acid microarray for detecting the ICAM-3 gene or a protein microarray for detecting the ICAM-3 protein.

In the microarray of the present invention, a primer, a probe or an antibody capable of measuring the expression level of the ICAM-3 protein or a gene encoding the ICAM-3 protein is used as a hybridizable array element and is immobilized on a substrate. Preferred substrates may include, for example, membranes, filters, chips, slides, wafers, fibers, magnetic beads or nonmagnetic beads, gels, tubing, plates, polymers, microparticles and capillaries, as suitable rigid or semi-rigid supports have. The hybridization array elements are arranged and immobilized on the substrate, and such immobilization can be performed by chemical bonding methods or covalent bonding methods such as UV. For example, the hybridization array element may be bonded to a glass surface modified to include an epoxy compound or an aldehyde group, and may also be bound by UV on a polylysine coating surface. In addition, the hybridization array element can be coupled to the substrate via a linker (e.g., ethylene glycol oligomer and diamine).

The "aptamer" means a nucleic acid oligomer having affinity for the ICAM-3 protein. The aptamer synthesizes randomly synthesized DNA oligomers having about 60 bases, separates the DNA oligomers binding to the ICAM-3 protein into affinity columns, amplifies and separates the ICAM-3 protein by repeating ICAM-3 Can be produced.

The anticancer drug resistance prediction kit of the present invention can be used for ICAM-3 gene detection or ICAM-3 protein detection.

When the anticancer drug resistance prediction kit of the present invention is used for gene amplification applied to the PCR amplification process, the kit of the present invention may optionally contain a reagent necessary for PCR amplification such as a buffer, a DNA polymerase (for example, Thermus aquaticus (Taq) Thermostable DNA polymerase obtained from Thermus thermophilus (Tth), Thermus filiformis, Thermis flavus, Thermococcus literalis or Pyrococcus furiosus (Pfu)), DNA polymerase joiner and dNTPs, and the anticancer drug resistance prediction kit Is for ICAM-3 protein detection applied in immunoassays, the kit of the present invention may optionally comprise a secondary antibody and a labeling substrate. Further, the kit according to the present invention may be manufactured from a number of separate packaging or compartments containing the reagent components described above.

Hereinafter, the present invention will be described in more detail with reference to examples.

[ Example ]

The specific methods used in this embodiment are described in detail in the above non-patent documents and in the Biochemical and Biophysical Research Communications 441 (2013) 507-513 published by the inventors of the present invention, Lt; / RTI >

1. Suppression of cell death ICAM -3 on anticancer drug resistance

3 was overexpressed in NCI-H1299, a non-small-cell cancer cell line, to construct a NCI-H1299 / ICAM-3 cell line (Fig. 3A) in order to observe whether ICAM- , The cell line was treated with anticancer drug by propidium iodide (PI) uptake method, and cell death was measured (Fig. 3B). As an anticancer agent, two kinds of paclitaxel and vincristine, which are anticancer drugs targeting the microtubules of cells, are used (Figs. 1A and 1B). These anticancer drugs are one of the anticancer drugs used in the treatment of lung cancer and are anticancer drugs that induce cell death by blocking the synthesis and degradation process of intracellular microtubules which actively grow like cancer cells. At 96 hours after treatment with anticancer drugs, NCI-H1299 / ICAM-3 cell line showed more than 30% inhibition of cell death, and PI uptake is a measure of cell death in various cell death mechanisms , And the change of intracellular apoptosis-related molecules after treatment with anticancer drug by Western blotting was measured (FIG. 3C). As shown in Fig. 3C, activation of caspase-3, 8, and 9 by the anticancer agent treatment was found to be less in the NCI-H1299 / ICAM-3 cell line than in the control group. In addition, ICAM-3 inhibits the activation of caspase-8 and caspase-9, the initiation caspases of the extrinsic pathway and the intrinsic pathway, It is also confirmed that the activation of caspase-3, Caspase-3, is also inhibited. In addition, activation of PARP, which appears as one of the indicator (indicator) of apoptosis (cleavage), too, and being inhibited by ICAM-3 overexpressed, wherein - Bcl-2, which has the features of apoptosis, Bcl-X _L, Mcl -1 was also inhibited by ICAM-3. In FIG. 3 D, cell death was observed by visual observation using a hematocytometer. As shown in the table, more than 30% reduction of cell death was observed in the overexpressed cell line of ICAM-3. In order to confirm that the inhibition of apoptosis by treatment with paclitaxel and vincristine was the main mechanism of inhibition of ICAM-3-induced apoptosis, apoptosis of zvAD-fmk, a pan-caspase inhibitor, Absorption method. After the inhibition of caspase activation by zVAD-fmk treatment, apoptosis was observed in both the control and ICAM-3 overexpressing cell lines. Therefore, it is thought that the apoptosis of paclitaxel and vincristine occurs through the signal transduction pathway in which apoptosis mainly occurs, and that the inhibition of apoptosis is thought to induce cell death by the anticancer agent, and ICAM -3 may also be inferred to result from suppression of the development of apoptosis.

2. ICAM -3 for anticancer drug resistance Akt  And ERK Activation effect of

Next, intracellular signal transduction mechanisms that act on the generation of anticancer drug resistance by ICAM-3 were studied. In this NCI-H1299 / ICAM-3 cell line, Western blotting was performed to observe representative tumor cell survival mechanism and activity and expression pattern of Akt and ERK molecules (FIG. 4A). As a result, it was observed that the NCI-H1299 / ICAM-3 cell line was continuously increased phosphorylation of Akt and ERK molecules. These results also indicate that Akt and ERK are indirectly involved in the down-signaling pathway of ICAM-3 in the intracellular signaling pathway of ICAM-3, as activation is stimulated with increased expression of ICAM-3. To investigate the effect of these signal transduction molecules, Akt and ERK, on the cytotoxic effect of paclitaxel and vincristine on cancer cell death, we used LY294002, an inhibitor of PI3 kinase, Akt, and PD98059, an inhibitor of ERK, After pretreatment, paclitaxel and vincristine were treated, and apoptosis was measured by the PI absorption method (Fig. 4B). As a result, cell death was significantly increased in the control and NCI-H1299 / ICAM-3 cell lines when the inhibitors LY294002 and PD98052 were treated, showing an increase rate of 50% or more. Therefore, activation of Akt and ERK molecules was found to be activated by the increase of anticancer drug resistance by ICAM-3. In addition, Western blotting was performed (FIG. 4C) to observe the increase of cell death by the anticancer drug when LY294002 and PD98052 were treated. The experimental results, the processing of each inhibitor was increased Paget CAS-3, 8, activation of 9, Bcl-2, the expression of Bcl-X _L, Mcl-1 confirmed that the significantly reduced. Therefore, it was observed that inhibition of the signal transduction pathway of Akt and ERK using the inhibitor increases the apoptosis inducing action of cancer cells by paclitaxel and vincristine. In addition, the inhibition of Akt and ERK signal transduction pathways in ICAM-3 over-expressing cell lines was comparatively low compared to the control group in Western blotting. In addition, cell counting assay was performed in combination with inhibitors and anticancer agents. The results of this experiment also showed that inhibition of Akt and ERK signaling pathway promotes more than 50% cell death. These results suggest that the increased expression of ICAM-3 induces an increase in Akt and ERK1 activity, which is important for anticancer drug resistance.

3. ICAM -3 for anticancer drug resistance CREB2  Effect of activation

The present inventors have confirmed that CREB2, which is one of the transcription factors, acts on the anticancer drug resistance-imparting action by ICAM-3 (FIG. 5). The relationship between ICAM-3 and the CREB family of transcription factor proteins has been demonstrated by the present inventors by demonstrating that increased activation of CREB1 by ICAM-3 increases the migration and invasiveness of cancer cells. In the present invention, , But it was not related. Instead, it was observed that the activity of CREB2 belonging to the CREB protein family was also increased by ICAM-3 (Fig. 5A), indicating that this increase in activity was related to increased Akt and ERK activity. Lt; RTI ID = 0.0 > ICAM-3 < / RTI > CREB2 expression inhibition assay using CREB2 siRNA showed that expression of CREB2 decreased during treatment with siRNA (Fig. 5C), and 30% or more of apoptosis was observed in the treatment with paclitaxel and vincristine after siRNA treatment with siRNA (Fig. 5D). In addition, we observed that the activity of caspase-8, 9 was increased when treated with CREB2 siRNA by Western blotting under the same conditions, and decreased expression of CREB2 and simultaneous treatment with anticancer drug induced cell death (FIG. 5E). Therefore, the results of this experiment suggest that CREB2 is located under the signaling pathway of ICAM-3-Akt / ERK and constitutes ICAM-3-Akt / ERK-CREB2 signal transduction, .

4. H460  Cancer cells and radiation In my castle ICAM -3 - Akt / ERK  - CREB2 Effect of anticancer drug resistance by activation

ICAM-3 was injected into the same non-small cell lung cancer cell line, NCI-H460 cell line, and NCI-H460 / NCI-H460 cells were transfected with the ICAM-3-Akt / ERK-CREB2 signal transduction- ICAM-3 cell line was established, and phosphorylation of Akt and ERK was also observed therein (FIG. 6A), confirming that CREB2 was also activated (FIG. 6B). 6). This cell line also showed a decrease in apoptosis when added with paclitaxel and vincristine (Fig. 6C), and this apoptosis inhibition was attributed to inhibition of apoptosis activity (Fig. 6D) . In addition to the non - small cell line, the radiation tolerance of the SiHa cell line, a cervical cancer cell, also confirmed the generation of anticancer drug resistance by activation of ICAM - 3 - Akt / ERK - CREB2 signaling. In the anticancer drug resistant strain SiHa / R, in which ICAM-3 was overexpressed, phosphorylation of Akt and ERK was increased (E in FIG. 6) and increased in activity of CREB2 (FIG. 6 ), And cell death was less in SiHa / R when treated with paclitaxel and vincristine (G in Fig. 6). In addition, it was observed that activation of apoptosis in SiHa / R occurs less in Western blotting (H in FIG. 6). Thus, it has been demonstrated that anti-cancer drug resistance is induced by activation of ICAM-3-Akt / ERK-CREB2 signaling in other non-small cell lung carcinomas other than NCI-H1299 as well as in radiation-tolerant ICAM-3 overexpressed.

5. Mouse Tumor  In the model ICAM -3 Effect of anticancer drug resistance

The cell death was less in the paclitaxel and vincristine injections of the xenografts constructed after the subcutaneous injection of the NCI-H460 / ICAM-3 cell line used in this experiment compared to the control group, (Fig. 7).

6. conclusion

As a result of the experiment conducted by the inventors of the present invention, it was confirmed that ICAM-3 is a radiation / anticancer drug cross-resistance inducing factor that induces not only radiation resistance but also anticancer drug resistance. In addition, the anticancer drug resistance by ICAM-3 was observed to be caused by inhibition of ICAM-3-Akt / ERK-CREB2 signaling-induced cell death, and activation of ICAM-3-Akt / ERK- And induction of anticancer drug tolerance by the inhibition of cell death is also found in various non - small cell lung cancer cell lines and radiation resistant strain. These results also confirmed that inhibition of cell death of ICAM-3 overexpressing cell lines was also observed in animal tests using mice. Therefore, it is expected that the development of inhibitors and therapeutic antibodies to ICAM-3 related signal transduction systems may lead to the development of new anti-cancer drugs. In summary, ICAM-3 has been shown to inhibit ICAM-3-Akt / CREB-1-MMP transactivation and invasion through the activation of FAK, the promotion of cancer cell proliferation through ERK, It has been proved through the present invention that it has an anticancer drug tolerance function through activation of ERK-CREB2 (FIG. 8).

<110> AAA <120> ICAM-3 AS BIOMARKER FOR THE PREDICTION OF TOLERANCE AGAINST          ANTICANCER AGENTS AND USE THEREOF <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 999 <212> DNA <213> Homo sapiens ICAM-3 nucleotide sequence <400> 1 atggccacca tggtaccatc cgtgttgtgg cccagggcct gctggactct gctggtctgc 60 tgtctgctga ccccaggtgt ccaggggcag gagttccttt tgcgggtgga gccccagaac 120 cctgtgctct ctgctggagg gtccctgttt gtgaactgca gtactgattg tcccagcttt 180 gagaaaatcg ccttggagac gtccctatca aaggagctgg tggccagtgg catgggctgg 240 gcagccttca atctcagcaa cgtgactggc aacagtcgga tcctctgctc agtgtactgc 300 aatggctccc agataacagg ctcctctaac atcaccgtgt acgggctccc ggagcgtgtg 360 gagctggcac ccctgcctcc ttggcagccg gtgggccaga acttcaccct gcgctgccaa 420 gtggagggtg ggtcgccccg gaccagcctc acggtggtgc tgcttcgctg ggaggaggag 480 ctgagccggc agcccgcagt ggaggagcca gcggaggtca ctgccactgt gctggccagc 540 agagacgacc acggagcccc tttctcatgc cgcacagaac tggacatgca gccccagggg 600 ctgggactgt tcgtgaacac ctcagccccc cgccagctcc gaacctttgt cctgcccgtg 660 acccccccgc gcctcgtggc cccccggttc ttggaggtgg aaacgtcgtg gccggtggac 720 tgcaccctag acgggctttt tccagcctca gaggcccagg tctacctggc gctgggggac 780 cagatgctga atgcgacagt catgaaccac ggggacacgc taacggccac agccacagcc 840 acggcgcgcg cggatcagga gggtgcccgg gagatcgtct gcaacgtgac cctagggggc 900 gagagacggg aggcccggga gaacttgacg gtctttagct tcctaggacc cattgtgaac 960 ctcagcgagc ccaccgccca tgaggggtcc acagtgacc 999 <210> 2 <211> 547 <212> PRT <213> Homo sapiens ICAM-3 amino acid sequence <400> 2 Met Ala Thr Met Val Pro Ser Val Leu Trp Pro Arg Ala Cys Trp Thr   1 5 10 15 Leu Leu Val Cys Cys Leu Leu Thr Pro Gly Val Gln Gly Gln Glu Phe              20 25 30 Leu Leu Arg Val Glu Pro Gln Asn Pro Val Leu Ser Ala Gly Gly Ser          35 40 45 Leu Phe Val Asn Cys Ser Thr Asp Cys Pro Ser Ser Glu Lys Ile Ala      50 55 60 Leu Glu Thr Ser Leu Ser Lys Glu Leu Val Ala Ser Gly Met Gly Trp  65 70 75 80 Ala Ala Phe Asn Leu Ser Asn Val Thr Gly Asn Ser Arg Ile Leu Cys                  85 90 95 Ser Val Tyr Cys Asn Gly Ser Gln Ile Thr Gly Ser Ser Asn Ile Thr             100 105 110 Val Tyr Gly Leu Pro Glu Arg Val Glu Leu Ala Pro Leu Pro Pro Trp         115 120 125 Gln Pro Val Gly Gln Asn Phe Thr Leu Arg Cys Gln Val Glu Gly Gly     130 135 140 Ser Pro Arg Thr Ser Leu Thr Val Val Leu Leu Arg Trp Glu Glu Glu 145 150 155 160 Leu Ser Arg Gln Pro Ala Val Glu Glu Pro Ala Glu Val Thr Ala Thr                 165 170 175 Val Leu Ala Ser Arg Asp His Gly Ala Pro Phe Ser Cys Arg Thr             180 185 190 Glu Leu Asp Met Gln Pro Gln Gly Leu Gly Leu Phe Val Asn Thr Ser         195 200 205 Ala Pro Arg Gln Leu Arg Thr Phe Val Leu Pro Val Thr Pro Pro Arg     210 215 220 Leu Val Ala Pro Arg Phe Leu Glu Val Glu Thr Ser Trp Pro Val Asp 225 230 235 240 Cys Thr Leu Asp Gly Leu Phe Pro Ala Ser Glu Ala Gln Val Tyr Leu                 245 250 255 Ala Leu Gly Asp Gln Met Leu Asn Ala Thr Val Met Asn His Gly Asp             260 265 270 Thr Leu Thr Ala Thr Ala Thr Ala Thr Ala Arg Ala Asp Gln Glu Gly         275 280 285 Ala Arg Glu Ile Val Cys Asn Val Thr Leu Gly Gly Glu Arg Arg Glu     290 295 300 Ala Arg Glu Asn Leu Thr Val Phe Ser Phe Leu Gly Pro Ile Val Asn 305 310 315 320 Leu Ser Glu Pro Thr Ala His Glu Gly Ser Thr Val Thr Val Ser Cys                 325 330 335 Met Ala Gly Ala Arg Val Gln Val Thr Leu Asp Gly Val Ala Ala             340 345 350 Ala Pro Gly Gln Pro Ala Gln Leu Gln Leu Asn Ala Thr Glu Ser Asp         355 360 365 Asp Gly Arg Ser Phe Phe Cys Ser Ala Thr Leu Glu Val Asp Gly Glu     370 375 380 Phe Leu His Arg Asn Ser Ser Val Gln Leu Arg Val Leu Tyr Gly Pro 385 390 395 400 Lys Ile Asp Arg Ala Thr Cys Pro Gln His Leu Lys Trp Lys Asp Lys                 405 410 415 Thr Arg His Val Leu Gln Cys Gln Ala Arg Gly Asn Pro Tyr Pro Glu             420 425 430 Leu Arg Cys Leu Lys Glu Gly Ser Ser Arg Glu Val Pro Val Gly Ile         435 440 445 Pro Phe Phe Val Asn Val Thr His Asn Gly Thr Tyr Gln Cys Gln Ala     450 455 460 Ser Ser Ser Gly Lys Tyr Thr Leu Val Val Val Met Asp Ile Glu 465 470 475 480 Ala Gly Ser Ser His Phe Val Pro Val Phe Val Ala Val Leu Leu Thr                 485 490 495 Leu Gly Val Val Thr Ile Val Leu Ala Leu Met Tyr Val Phe Arg Glu             500 505 510 His Gln Arg Ser Gly Ser Tyr His Val Arg Glu Glu Ser Thr Tyr Leu         515 520 525 Pro Leu Thr Ser Met Gln Pro Thr Glu Ala Met Gly Glu Glu Pro Ser     530 535 540 Arg Ala Glu 545

Claims (13)

The expression levels of ICAM-3 (intercellular adhesion molecule-3) in cancer cells isolated from non-small cell lung cancer or cervical cancer patients treated with paclitaxel or vincristine-specific anticancer drugs were measured, and cancer cells Which is judged to have a possibility of resistance to an anticancer agent when the expression amount of ICAM-3 measured from cancer cells isolated from a cancer patient after treatment with an anticancer drug is increased as compared with the amount of ICAM-3 detected from a cancer patient A method of providing information for anticancer drug resistance prediction of paclitaxel or vincristine in a cell lung cancer or cervical cancer. The method of claim 1, wherein the method comprises the steps of:
A first step of preparing a sample from cancer cell tissue isolated from non-small cell lung cancer or cervical cancer patients and measuring the amount of ICAM-3 expression from the sample;
A second step of preparing a sample from cancer cell tissue isolated from non-small cell lung cancer or cervical cancer patients to which an anti-cancer agent specified by paclitaxel or vincristine is administered, and measuring the amount of ICAM-3 expression from the sample;
Comparing the result of the first step with the result of the second step;
Providing information for determining that the possibility of resistance to an anticancer drug is high when the amount of expression of ICAM-3 in the second step is higher than the amount of expression of ICAM-3 in the first step through the comparison. delete delete 3. The method according to claim 1 or 2, wherein the measurement of the expression level of ICAM-3 measures the amount of mRNA or protein. 6. The method according to claim 5, wherein the measurement of the amount of mRNA is performed by reverse transcriptase-polymerase chain reaction (RT-PCR), northern blotting, or nucleic acid microarray. 6. The method according to claim 5, wherein the measurement of the amount of protein is a measurement by an antigen-antibody reaction. The method according to claim 7, wherein the measurement by the antigen-antibody reaction is detection by Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry or protein microarray or aptamer Lt; / RTI &gt; delete delete delete A kit for predicting anticancer drug resistance specific to paclitaxel or vincristine in non-small cell lung cancer or cervical cancer, comprising means for measuring the expression level of intercellular adhesion molecule-3 (ICAM-3). 13. The kit of claim 12, wherein the means is selected from the group consisting of primers, probes, antibodies, antibody fragments, microarrays and aptamers.

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