KR101326367B1 - Diagnostic methods and kits for monitoring resistance to therapeutic chemoagent - Google Patents

Abstract

The present invention relates to a diagnostic method and a diagnostic kit for anticancer drug resistance, and more particularly, a gene for predicting drug resistance to drugs for treating chronic myelogenous leukemia (CML), and anticancer drug resistance using the same. Relates to diagnostic methods and diagnostic kits. The diagnostic method and diagnostic kit of the present invention detects gene expression signals that impart resistance to imatinib, an anticancer agent, using various resistance gene groups, and effectively diagnoses and treats various cancers including chronic myelogenous leukemia. Furthermore, it can be widely used to study the mechanism of resistance to Gleevec.

Anticancer drug resistance, imatinib, resistance gene family, chronic myelogenous leukemia (CML), diagnostic kit

Description

Diagnostic methods and kits for monitoring resistance to therapeutic chemoagent

1 shows the sequence of imatinib-resistant gene groups according to gene expression profiles.

FIG. 2A analyzes the expression change of positively correlated genes according to the dose of imatinib using imatinib-resistant gene group.

FIG. 2B analyzes the expression changes of negatively correlated genes according to the dose of imatinib using imatinib-resistant gene group.

FIG. 3A shows CASP4 resistance among six candidate genes for imatinib resistance in chronic myelogenous leukemia (CML). And TNFRSF21 It is analyzed by the change of expression of gene.

3B shows RAB11A resistance among imatinib resistance of six candidate genes in chronic myelogenous leukemia (CML) And IGF1 Figure 3c shows the LYN of imatinib resistance in chronic myeloid leukemia (CML) among six candidate genes. And RHOA It is analyzed by the change of expression of gene.

The present invention relates to a diagnostic method and a diagnostic kit for anticancer drug resistance, and more particularly, a gene for predicting resistance to an agent for treating chronic myelogenous leukemia (hereinafter, abbreviated as? CML?). The present invention relates to a diagnostic method and a diagnostic kit for anticancer drug resistance.

The anticancer agent imatinib is a selective inhibitor of tyrosine kinase, called imatinib mesylate, STI-571, and Gleevec, and is known as chronic myelogenous leukemia; Abbreviated? Imatinib, the primary drug for treating BCR-ABL fusion kinase, is known to alleviate more than 80% of symptoms, especially in the early stages of chronic myeloid leukemia (O'Dwyer, ME and Druker, BJ J. Intern. Med . 250: 3-9, 2001) . However, symptoms recur frequently after initial blood and cellular responses (Druker, BJ et al ., N. Engl . J. Med . , 344: 1038-1042, 2001). Thus, therapies targeting BCR-ABL kinase are hampered by drug resistance and severely reduce the clinical efficacy of imatinib.

In addition, BCR-ABL amplification transcription (transcription amplification) or point mutations in the constant domain of a tyrosine kinase is often been observed in other clinical cases of resistance or in vivo (in vitro) cell model (Hochhaus, A. et al., Science , 293: 876-880, 2001; Shah, NP et al. , Cancer Cell , 2: 117-125, 2002; Gorre, ME et al ., Science , 293: 876-880, 2001). Although this change is the most likely cause of obtaining imatinib-resistance, previous studies have shown that this resistance can occur without amplification or mutation (Donato, NJ et al ., Blood , 101: 690-698, 2003; Hofmann, WK et al ., Lancet, 359: 481-486, 2002; Mahon, FX et al ., Blood, 96: 1070-1079, 2000). In addition, overexpression of a multidrug resistant gene also confers resistance to imatinib. These studies suggest that imatinib-resistance can be caused by different mechanisms. Thus, a variety of approaches have been introduced, such as general purpose gene expression analysis using microarrays to identify the major factors causing imatinib-resistance (Tipping, AJ et al. , Exp . Hematol . , 31: 1073-1080 , 2003; Ohmine, K. et al. , Stem Cells , 21: 315-321, 2003).

We established an imatinib-resistant gene family with reference to various imatinib doses and CML cases and examined imatinib-resistant expression profiles to focus on imatinib-resistant transcripts showing biologically appropriate expression changes. The gene expression signature associated with the gene was identified. In addition, bioinformatics approaches such as gene set enrichment analysis attempted to describe the expression profiles and candidate signaling processes associated with imatinib-resistance.

Therefore, the present inventors have continued to develop a diagnostic method for testing new anticancer drug resistance, and as a result, the gene signal conferring resistance to imatinib, an anticancer drug treating chronic myelogenous leukemia (CML) The present invention was successfully completed by selecting 6 isolates of resistance genes, producing various gene probes, and developing a diagnostic kit, and then confirming the usefulness of testing drug resistance of various cancers including CML.

An object of the present invention is to provide an anticancer drug resistance gene, a method for diagnosing anticancer drug resistance and a diagnostic kit using the same.

In order to achieve the above object,

(1) selecting anticancer drug resistance genes; And

(2) analyzing the results by measuring the expression level of the gene; It provides a diagnostic method for testing resistance to the anticancer drug.

In the process (1), the anticancer drug resistance gene is preferably at least one selected from apoptosis, cell signal transduction, cell cycle regulation, GTP binding protein, cell proliferation and transcription related genes.

The anticancer agent resistance gene is CASP4, TNFRSF21, TNFRSF9, NOTCH2, SQSTM1, TGIF, PLCD1, CCNB1P1, RAB11A, IGF1, LYN, RHOA, NMI, FOXO3A, ZNF226, ZNF140, LTBP1 and ABCA8 gene preferably at least one selected from a, and CASP4 More preferably, at least one selected from among TNFRSF21, RAB11A, IGF1, LYN and RHOA genes.

In the process (2), the expression level of the anticancer drug resistance gene is increased or decreased, and at this time, the CASP4, TNFRSF21, LYN and / or RHOA genes are up-expressed and the RAB11A and / or IGF1 genes are decreased.

In the step (2), the expression level of the anticancer drug resistance gene is preferably analyzed by performing polymerase chain reaction (PCR) and / or microarray.

The diagnostic method of the present invention is preferably used to treat chronic myeloid leukemia (CML).

In addition, the present invention provides a marker for an anticancer agent resistance gene selected from apoptosis, cell signal transduction, cell cycle regulation, GTP binding protein, cell proliferation and transcription related genes and all or a portion thereof. . The anticancer agent resistance gene is CASP4, TNFRSF21, TNFRSF9, NOTCH2, SQSTM1, TGIF, PLCD1, CCNB1P1, RAB11A, IGF1, LYN, RHOA, NMI, FOXO3A, ZNF226, ZNF140, LTBP1 and ABCA8 gene preferably at least one selected from a, and CASP4 More preferably, at least one selected from among TNFRSF21, RAB11A, IGF1, LYN, and RHOA genes, and is preferably used to treat chronic myeloid leukemia (CML).

Hereinafter, the present invention will be described in detail.

The present invention provides a diagnostic method for testing resistance to anticancer drugs using a probe for anticancer drug resistance genes.

The anticancer agents include imatinib, Gleevec and STI-571. In addition, the anticancer drug resistance gene includes one or more selected from the CASP4, TNFRSF21, RAB11A, IGF1, LYN and RHOA gene.

The diagnostic method of the present invention may include any test method using a probe for an anticancer drug resistance gene, and preferably include a polymerase chain reaction performed using the probe for the resistance gene. More preferably the anticancer drug resistance gene CASP4 A primer set having a nucleotide sequence of SEQ ID NO: 1 and SEQ ID NO: 2 for;

Anticancer drug resistance gene TNFRSF21 A primer set having a nucleotide sequence of SEQ ID NO: 3 and SEQ ID NO: 4 for a;

Anticancer drug resistance gene RAB11A A primer set having a nucleotide sequence of SEQ ID NO: 5 and SEQ ID NO: 6 for a;

Anticancer drug resistance gene IGF1 A primer set having the nucleotide sequences of SEQ ID NO: 7 and SEQ ID NO: 8 for the;

A primer set having a nucleotide sequence of SEQ ID NO: 9 or SEQ ID NO: 10 for an anticancer drug resistance gene LYN ; And

Provided is a set of anti-cancer drug resistance diagnostic primers, characterized in that at least one selected from the group consisting of a primer set having the nucleotide sequences of SEQ ID NO: 11 and SEQ ID NO: 12 for the anti-cancer drug resistance gene RHOA. In addition, the present invention provides a method for diagnosing resistance to cancer, wherein the polymerase chain reaction (PCR) is performed using the primer set. The diagnostic method of the present invention is preferably used to treat various cancer diseases including chronic myeloid leukemia (CML).

The present invention provides a diagnostic kit for testing resistance to an anticancer agent using a probe for an anticancer agent resistance gene. Preferably, a diagnostic kit for testing resistance to an anticancer agent comprising a nucleic acid sequence encoding at least one gene selected from an anticancer agent resistance gene CASP4, TNFRSF21 , RAB11A, IGF1, LYN, and RHOA is provided. More preferably, it further comprises a nucleic acid sequence encoding at least one gene selected from TNFRSF9 , NOTCH2 , SQSTM1 , TGIF , PLCD1 , CCNB1P1 , NMI , FOXO3A , ZNF226 , ZNF140 , LTBP1 and ABCA8 genes in addition to the anticancer drug resistance gene. A diagnostic kit for testing resistance to an anticancer agent is provided. The anticancer drug resistance diagnostic kit can be used to treat chronic myeloid leukemia (CML). The anticancer agents include imatinib, Gleevec and STI-571. In addition, the anticancer drug resistance gene includes one or more selected from the CASP4, TNFRSF21, RAB11A, IGF1, LYN and RHOA gene. The diagnostic kit of the present invention is preferably used to treat various cancer diseases including chronic myeloid leukemia (CML).

The diagnostic kit of the present invention may include all test reagents and devices using probes for anticancer drug resistance genes, and preferably reagents for performing polymerase chain reaction using the probes for the resistance genes. And devices.

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

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Example  1> Imatinib - Resistance K562  Examining Expression Profiles of Cell Lines

First, in the present invention, in order to establish an imatinib-resistant cell line, resistance to imatinib was induced by increasing the concentration of imatinib by 50 mM every two weeks in a leukemia K562 cell line that becomes a red blood cell. During the process the imatinib culturing cells imatinib various concentrations 200 nM, 400 nM, 600 nM resistant cell lines and four series, each exposed to 800 nM, _200nM KR, KR _{400nM, 600nM} and KR KR _800nM Was established. Mutations in the BCR-ABL kinase domain at this time were detected as previously described (Shah, NP et al. , Cancer Cell , 2: 117-125, 2002). In addition, the expression level of the BCR-ABL kinase domain was measured by performing real time reverse transcriptase-polymerase chain reaction (RT-PCR) using primers used for mutation analysis. In this case, human GAPDH was used as a control.

First, it was investigated whether the imatinib resistance of these cell lines was due to increased transcriptional activity, as previously reported in previous studies, or whether it was caused by point mutations in the BCR-ABL kinase domain (Shah, NP et al ., Cancer Cell , 2). : 117-125, 2002). However, neither point mutations in the specific region of the kinase domain nor synergistic transcriptional regulation of the BCR-ABL gene were observed in four resistant cell lines. Expression profiles of K562 cell lines and imatinib resistant cell lines were measured using oligo-microarrays containing as many as 30,000 human genes. The population of 3,000 most diverse transcripts allowed cell lines to be distinguished from each other by imatinib doses that induce resistance (see FIG. 1). According to such grouping, struggled yeoryang group, i.e. KR KR _600nM and _800nM are low dose group, i.e. KR KR _200nM and _400nM And it was confirmed that it is clearly distinguished from K562 parental cells.

Example  2. In anticancer drug resistant cell lines Transcripts  Change analysis

(1) RNA isolation and Hybridization

First, total RNAs were isolated from K562 parental and imatinib-resistant cell lines using Trizol (Invitrogen) according to the manufacturer's manual. The quantity and quality of the isolated RNAs were assessed using a Bioanalyzer (Bioanalyzer 2100; manufactured by Agilent Technologies). Human Genome Survey Microarray (applied by Applied Biosystems) version 2.0 containing about 30,000 human gene sets was used to express the expression profiles of five cell lines, namely sensitive K562 cells and four resistant cell lines. Analyzed. CRNA labeled with deoxygenin-UTP was made and in turn amplified from 5 μg total RNAs using the chemifluorescence RT-IVT labeling kit version 2.0 (Applied Biosystems). Array hybridization, chemifluorescence testing and imaging and analysis were performed according to the manufacturer's protocol using a chemifluorescence kit and a chemifluorescence microarray analyzer 1700 (Applied Biosystems). The hybridization reaction was repeated three times for each sample.

(2) Data  Processing

Images of each array-hybridization were collected using a 1700 analyzer equipped with a high-resolution optical format CCD camera. This includes two chemifluorescent images exposed for 5 seconds to analyze gene expression, two fluorescent images for feature studies and spot normalization, and two QC images for spectral correction. These images were automatically entered (gridded) to quantify the chemifluorescent signal. In light of the background, the intensity data of the spots were adjusted using the deviation stabilization and normalization (VSN) method (Huber, W. et al. , Bioinformatics , 18: S96-104, 2002).

(3) differentially expressed Transcripts  analysis

Average expression of 29,098 genes from five cell lines, including K562 blasts, was assessed by one-way ANOVA analysis. The 3,000 mutants with the most variation were selected, and populations with average associations were formed using Cluster and Treeview ( http://rana.lbl.gov/EisenSoftware.htm ) software. To investigate whether the transcripts are synergistically or diminishedly regulated in resistant cell lines, expression changes were evaluated by comparing the relative blast control and calculating relative differences.

(4) Investigation of Dose-dependent Expression Changes

In order to identify genes whose expression changes in dose-resistant cell lines, two statistical approaches were combined (Tseng, YH et al. , Nat. Cell Biol . , 7: 601-611, 2005). First, five average expression levels were considered consecutively in the control and four resistant cell lines, with imatinib doses ranging from 200 nM to 800 nM. The importance of synergistic and declining progression was determined according to the hypothesis of Equation 1 or the inverse of Equation 2.

First, as a result of investigating synergistically or diminishingly regulated transcripts in all four cell lines resistant to K562 cells used as a control, it was observed that commonly changed transcripts are over or underexpressed (Table 1). Reference). Specifically, apoptosis related genes, namely CASP4 , TNFRSF21 and TNFRSF9, are commonly upregulated in resistant cell lines, and also genes associated with transcription and signaling, such as NOTCH2 , SQSTM1 and TGIF, are reduced.

Especially NOTCH2 The gene is known to be a positive regulator of RAS signal transduction in its product, and is involved in molecular interconnection with mitogen-activated protein (MAP) kinases (Aster, JC and Pear, WS). . Curr Opin Hematol, 8: 237-244 , 2001; Zeng, Q. et al, Cancer Cell, 8:... 13-23, 2005). TGIF is also a co-repressor that forms negative feedback in TGF-β signaling. Among the expression decreasing genes, IGF1 and PLCD1 genes are involved in signal transduction, and IGF1 has been reported to encode growth factors that regulate somatic growth and development as well as carcinogenesis (Zumkeller, W., Mol . Pathol ., 54: 285-288, 2001). In addition, the PLCD1 gene produced the second messenger molecules diacylglycerol and inositol-1,4,5-triphosphate and was involved in signal transduction (Stallings, JD et al. , J. Biol . Chem . , 280: 22060-22069, 2005). In addition, molecules related to the cell cycle such as CCNB1P1 have been identified and reported to act on the progression of the cell cycle through the G ₂ / M phase (Toby, GG et al. , Mol . Cell. Biol . , 23: 2109- 2122, 2003).

Example  3. Imatinib In resistant cell lines Imatinib  Investigate changes in expression related to dosage

Genes showing dose-response between gene expression and imatinib doses were examined in four resistant cell lines (see Table 2). Figure 2 is a change analysis of gene expression according to the dose of imatinib using imatinib-resistant gene group. Of the 22 genes correlated with an increase in imatinib dosage, four genes involved in transcription, NMI , FOXO3A , ZNF226 And ZNF140 Was confirmed.

Specifically, proteins encoded from the NMI gene react with electron regulators and electron activators (STATs), such as N-MYC, C-MYC and signal transducers. In addition, proteins encoded from the NMI gene were found to be highly expressed in myeloid leukemia (Bao, J. and Zervos, AS, Oncogene , 12: 2171-2176, 1996). LTBP1, a transcript involved in signal transduction, is also involved in the TGF-β signaling process and produces proteins that react with TGF-β molecules. ABCA8, an ATP-dependent drug transporter, has also been identified among genes with positive expression changes associated with imatinib doses (Tsuruoka, S. et al., Biochem . Biophys . Res. Commun . , 298: 41- 45, 2002). Genes associated with cell proliferation have been identified among transcripts that negatively correlated with imatinib dose. In addition, translocation of MLF1 has often been observed in myelodysplastic syndromes and acute myeloid leukemia (Matsumoto, N. et al. , Leukemia , 14: 1757-1765, 2000). Among the genes involved in metabolism and signal transduction, RAB11A is a GTP binding protein associated with RAS that acts on carcinogenesis.

Example  4. Gene Set Amplification Analysis

Functionally related genes: GO (Gene Ontology), BioCarta ( http://www.biocarta.com ) and KEGTO (Kyoto Encyclopedia of Genes and Genomes), and Gene MicroArray Pathway Profiler (GenMAPP) were collected from the general genetic database ( Harris, MA et al. , Nucleic Acids Res. , 32: D258-D261, 2004; Kanehisa, M. et al ., Nucleic Acids Res. , 30: 42-46, 2002; Dahlquist, KD et al ., Nat. Genet. , 31: 19-20, 2002). Genes were matched and sorted into sets of genes with specific markers, and 3,774 sets of genes with common functional markers but not mutually exclusive were collected. Amplification analysis of genes was done using exponential analysis (PAGE), which directly calculates importance based on Z-statistics. To perform the analysis, initial gene sets were limited to 721 sets containing at least 10 genes corresponding to the minimum gene set. In this case, two kinds of index values, ie, relative differences rd and F _k, are used to calculate the importance based on Z-statistics.

As a result of measuring and analyzing the statistical significance of amplification for gene sets of about 700 known functions in imatinib-resistant gene sets, the gene sets activated or suppressed were identified as shown in Table 3.

The 15 activated gene sets are as follows: four gene sets involved in transcriptional regulation; Three sets of genes that appear to be involved in transcription such as DNA binding, nucleic acid binding, and zinc ion binding; Two sets of genes associated with apoptosis; And a set of genes associated with the TGF-β signaling process. In addition, the seven suppressed gene sets are: gene sets involved in biosynthesis, transport and folding and protein metabolism; And three gene sets associated with small GTPase signaling.

Example  5. Myeloid Leukemia In patients  Gene Expression Profile Investigation

In order to investigate the gene expression profile of CML patients, the present invention was performed in real time quantitative PCR analysis as follows. Total RNA from blood samples from six CML patients before and after imatinib resistance was obtained was used for real-time QPCR analysis for six candidate genes: CASP4, TNFRSF21, RAB11A, IGF1, LYN and RHOA . As a result, three of the patients had a BCR-ABL mutation and no mutations were found in the other three. The first cDNA strand was synthesized using M-MLV reverse transcriptase (Invitrogen) and used for PCR. Real-time QPCR was performed using the Mx300P QPCR system with MxPro version 3.00 software from Stratagene. 20 μL of real time QPCR reaction was added with 10 ng cDNA, 1 × SYBR Green Tbr polymerase mixture, 0.5 × ROX and 20 pmole primer. In the present invention, a primer comprising a nucleotide sequence of SEQ ID NO: 1 to SEQ ID NO: 14 was synthesized and used. GAPDH was used as a control for each process.

The QPCR procedure was: 95 ° C., 10 minutes; 94 ° C., 10 seconds, 40 repetitions; 55-60 ° C., 30 seconds; And 72 ° C., 30 seconds. In order to confirm the specific amplification, the melting (melting curve) analysis was performed at 55-95 ℃, 0.5 ℃ / sec. Relative quantification was performed by the ΔC _t method.

Six CML patients with imatinib resistance were tested to determine whether the expression changes resulting from obtaining imatinib resistance in an in vitro model were actually appropriate for leukemia (CML) patients. The log ratio of expression change (Log ₂ ratio) before and after obtaining the resistance was calculated for each patient depending on the presence of the BCR-ABL mutation (n = 3) (see FIG. 3). 3A, 3B and 3C show the imatinib resistance of chronic myeloid leukemia (CML) as a change in expression of six candidate genes. At this time, the expression ratio between K562 and KR _800nM was the reference.

Specifically, CASP4 and TNFRSF21 were synergistically regulated in four and three patients after obtaining imatinib resistance, but this expression rate was not clearly distinguished between positive and negative patients for BCR-ABL mutations. On the other hand, RAB11A and IGF1 were consistently reduced in BCR-ABL mutant negative patients and elevated in 2 of 3 BCR-ABL mutant positive patients. In addition, LYN and RHOA which have already been reported to be elevated in relation to imatinib resistance Was also investigated. Although they are increased in cell lines without BCR-ABL mutations, this study was found to be underexpressed in two of the three mutant negative patients. In addition, expression of LYN and RHOA was significantly elevated in two out of three mutant positive patients.

As described above, the present invention relates to a gene for predicting drug resistance for treating chronic myelogenous leukemia (CML), a diagnostic method and a diagnostic kit for anticancer drug resistance using the same, and an anticancer agent imatinib A gene expression signal that confers resistance to imatinib is detected using various groups of resistance genes, and is widely used to effectively diagnose and treat various cancers including chronic myelogenous leukemia and further study the mechanism of resistance to Gleevec. Can be used.

<110> CATHOLIC UNIVERSITY INDUSTRY ACADEMIC COOPERATION FOUNDATION <120> Diagnostic methods and kits for monitoring resistance to          therapeutic chemoagent <130> PN060075 <160> 14 <170> Kopatentin 1.71 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CASP4-Forward primer <400> 1 acacgcctgg ctctcatcat a 21 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> CASP-4 Reverse primer <400> 2 cacagttccc cacggtttg 19 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> TNFRSF21 Foward Primer <400> 3 caaggccccc accacagaca catc 24 <210> 4 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> TNFRSF21 Reverse Primer <400> 4 gccggggccc ctttttcaga gt 22 <210> 5 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> RAB11A Forward primer <400> 5 acccgcgacg acgagta 17 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> RAB11A Revers Primer <400> 6 gcccacaagc atgataacaa t 21 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IGF1 Forward Primer <400> 7 gcggggctga gctggtggat 20 <210> 8 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> IGF1 Reverse Primer <400> 8 ggtcttgggc atgtcggtgt gg 22 <210> 9 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> LYN Forward Primer <400> 9 atacatcgag cggaagaact acat 24 <210> 10 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> LYN Reverse Primer <400> 10 acagggcggt catcacg 17 <210> 11 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> RHOA Forward Primer <400> 11 acagctgggc aggaagatta tgat 24 <210> 12 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> RHOA Reverse Primer <400> 12 acaagacaag gcacccagat tttt 24 <210> 13 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> GAPDH Forward Primer <400> 13 gcggggctct ccagaacatc at 22 <210> 14 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH Reverse Primer <400> 14 ccagccccag cgtcaaaggt g 21

Claims (9)

A diagnostic kit for testing resistance to an anticancer agent comprising all nucleic acid sequences encoding the anticancer agent resistance genes CASP4, TNFRSF21, RAB11A, IGF1, LYN, and RHOA, respectively. delete The method of claim 1, The kit is a diagnostic kit for testing resistance to anticancer drugs, characterized in that used to treat chronic myelogenous leukemia (CML). A method for providing information necessary for diagnosing resistance to an anticancer agent comprising measuring all expression levels of each of the anticancer agent genes CASP4, TNFRSF21, RAB11A, IGF1, LYN, and RHOA . delete 5. The method of claim 4, The method is used to treat chronic myeloid leukemia (CML). 5. The method of claim 4, The gene expression level is analyzed by performing polymerase chain reaction (PCR) or microarray (microarray) method for providing information necessary for diagnosing resistance to the anticancer agent. A primer set having the nucleotide sequences of SEQ ID NO: 1 and SEQ ID NO: 2 for the anticancer drug resistance gene CASP4 ; A primer set having a nucleotide sequence of SEQ ID NO: 3 and SEQ ID NO: 4 for an anticancer drug resistance gene TNFRSF21 ; A primer set having the nucleotide sequences of SEQ ID NO: 5 and SEQ ID NO: 6 for the anticancer drug resistance gene RAB11A ; A primer set having the nucleotide sequences of SEQ ID NO: 7 and SEQ ID NO: 8 for the anticancer drug resistance gene IGF1 ; A primer set having a nucleotide sequence of SEQ ID NO: 9 or SEQ ID NO: 10 for an anticancer drug resistance gene LYN ; And An anticancer drug resistance diagnostic primer set comprising both a primer set having the nucleotide sequences of SEQ ID NO: 11 and SEQ ID NO: 12 for an anticancer drug resistance gene RHOA. The method of claim 7, wherein the polymerase chain reaction (PCR) is performed using the primer set according to claim 8.

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