JP2010268690A - Method for determining sensitivity of compound having anti-tumor activity - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for simply and surely determining the sensitivity of an anti-cancer agent of AhR agonist, because it has been impossible to preliminarily predict the sensitivity of the anti-cancer agent of AhR agonist, although it has been found that cancers to which the anti-cancer agent of AhR agonist is effective and cancers to which the anti-cancer agent of AhR agonist is not effective are clearly divided. <P>SOLUTION: There is provided the method for clearly distinguishing cancers having sensitivity to the anti-cancer agent of AhR agonist and cancers not having the sensitivity by hierarchical cluster analyses using 266 genes each having a significant gene expression level difference (P<1.0×10<SP>-5</SP>) between cells having sensitivity and cells not sensitivity to the anti-cancer agent of AhR agonist, thereby predicting the therapeutic effect of the compound. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method for determining sensitivity using a gene set for predicting the sensitivity of a primary cancer tissue to a compound exhibiting antitumor activity. The determination method of the present invention is useful when determining the therapeutic effect of the above-mentioned compound or derivative thereof or pharmacologically acceptable salt thereof for cancer patients.

  The number of female breast cancer cases in Japan has increased remarkably, and in 1998, it surpassed gastric cancer and became the top female cancer patient. The number of affected cases is expected to increase to 41500 in 2005, 45600 in 2010, and 48500 in 2015. Drug therapy for breast cancer includes preoperative chemotherapy, postoperative adjuvant therapy, and drug therapy for recurrent breast cancer. Among them, estrogen receptor positive [ER (+)] patients are indicated for antihormonal treatment, but only half of them (about 30% of all breast cancer patients) are effective, so they have a wider spectrum. A breast cancer drug was desired.

  Under such circumstances, (5S, 7S) -7-methyl-3- (3-trifluoromethylphenyl) -5,6,7,8-tetrahydrocinnolin-5-ol (hereinafter referred to as 150460) It has become clear that ER (+) breast cancer cells that do not respond to existing endocrine therapy agents, breast cancer cells that have relapsed with anti-estrogen treatment, and a broad spectrum of antitumor effects against some ER (-) breast cancer cells. (Patent Document 1). Further, a prodrug (hereinafter referred to as 150460-P) in which solubility is improved by adding succinic acid to 150460 has been developed (Patent Document 2). This prodrug is presumed to produce 150460 by the action of hydrolase in vivo.

  2- (4-Amino-3-methylphenyl) -5-fluorobenzothiazole (hereinafter referred to as 5F-203) has been found as a compound showing an antitumor spectrum similar to 150460 (FIG. 1). 5F-203 acts as an allyl hydrocarbon receptor agonist (hereinafter referred to as AhR agonist) and induces CYP1A1, which is a downstream gene. Furthermore, it has been reported that the CYP1A1 activity is necessary for the antitumor effect (Non-patent Document 1, Non-patent Document 2). Although the molecular species of CYP is expressed in various tissues, it is often expressed particularly in the liver. As a role of this, the low molecule that has entered the body is hydroxylated to increase the hydrophilicity, thereby promoting extracorporeal elimination of foreign substances. It is considered to be. On the other hand, it is generally known that it contributes to the medicinal effect by participating in the activation of some drugs. In addition, (5-amino-2,3-fluorophenyl) -6,8-difluoro-7-methyl-4H-1-benzopyran-4-one (hereinafter referred to as NSC686288) is a compound that acts by the same mechanism as 5F-203. Etc.) have also been found (Non-Patent Document 3). Furthermore, molecular species such as CYP1A2 and CYP1B1 exist downstream of AhR, and are known to be involved in drug metabolism. Therefore, a compound that acts as an AhR agonist and is activated by CYP1A2 or CYP1B1 to exhibit an antitumor effect is considered to act on cells similar to those in which 150460, 5F-203, and NSC686288 are effective.

  For 5F-203, expression of each gene in the SAGE library using BG1, ZR-75-1 and MCF-7 which are three types of sensitive breast cancers, and Hs0578T and MDA-MB-231 which are non-sensitive breast cancers. A report has been made on genes having a significant difference in expression level between sensitive cells and non-sensitive cells analyzed by frequency and confirmed by quantitative PCR (Non-patent Document 4). However, the effect of an anticancer agent having an AhR agonist action cannot be reliably predicted.

International Publication No. 2004/052866 Pamphlet International Publication No. 2005/121105 Pamphlet International Publication No. 2001/014354 Pamphlet International Publication No. 1996/024592 Pamphlet Loaiza-P <e> rez AI, Trapani V et al. Mol. Pharmacol. 61 (1): 13-19 (2002) Loaiza-P <e> rez AI, Kenney S et al. Mol. Cancer Ther. 3 (6): 715-725 (2004) Kuffel MJ, Schroeder JC, Pobst LJ et al. Mol. Pharmacol. 62 (1): 143-153 (2002) Wallqvist A, Connelly J et al. Mol. Pharmacol. 69 (3): 737-748. (2006)

  When these anti-cancer agents, which are AhR agonists, are administered to patients, there is no method for determining the presence or absence of sensitivity before drug administration, which has been a big problem. From the study of cultured cancer cell lines, the anti-cancer drugs that are AhR agonist are clearly separated into cells that do not show the effect and cells that do not show (Fig. 1). Establishment was strongly desired.

  An object of the present invention is to provide a simple and reliable method for determining the sensitivity of cancer cells to an anticancer agent which is AhR agonist. That is, it is to provide a method for predicting whether or not an anticancer agent which is AhR agonist is effective for treatment.

  The inventors of the present application have found that by measuring the expression level of a specific gene in a cancer cell, the presence or absence of sensitivity to a cancer cell of a compound having AhR agonist and antitumor activity can be clearly distinguished. Further, it was proved that sensitivity could be determined by performing cluster analysis of nude transplanted subcutaneous tumor derived from human breast cancer whose sensitivity of 150460 is unknown, and the present invention was completed.

That is, the present invention
(1) AhR agonist and a compound exhibiting antitumor activity, comprising a step of measuring the expression level of one or more genes selected from a gene group consisting of 266 genes represented by SEQ ID NOs: 1 to 266, A method for determining the sensitivity of a cancer cell to a derivative, or a pharmacologically acceptable salt thereof,
(2) The compound is (5S, 7S) -7-methyl-3- (3-trifluoromethylphenyl) -5,6,7,8-tetrahydrocinnolin-5-ol, 2- (4-amino-3) -Methylphenyl) -5-fluorobenzothiazole, (5-amino-2,3-fluorophenyl) -6,8-difluoro-7-methyl-4H-1-benzopyran-4-one or their pharmacologically The determination method according to the above (1), which is an acceptable salt thereof,
(3) The AhR agonist described in (1) or (2) above, wherein the gene group is a gene group consisting of 25 genes represented by SEQ ID NOs: 1 to 25, a compound exhibiting antitumor activity, a derivative thereof, Or a method for determining the sensitivity of a cancer cell to a pharmacologically acceptable salt thereof,
(4) The method according to any one of (1) to (3), wherein the cancer cells are breast cancer cells, liver cancer cells, or ovarian cancer cells,
(5) An AhR agonist having anti-tumor activity to which an oligo DNA complementary to the full length of one or more genes selected from the group consisting of 25 genes described in (3) above or a partial sequence thereof is fixed. A microarray for determining the sensitivity of cancer cells to a compound, a derivative thereof, or a pharmacologically acceptable salt thereof,
(6) A kit for use in a method for determining sensitivity to a compound or derivative thereof having antitumor activity, or a pharmacologically acceptable salt thereof, comprising the microarray according to (5).
(7) A primer set for measuring the expression level of one or more genes selected from the gene group described in (1) above,
(8) An antibody set for quantifying the amount of intracellular protein of one or more genes selected from the gene group described in (1) above,
About.

  By using mRNA extracted from cancer tissue isolated from a cancer patient and comparing the gene expression pattern between sensitive and non-sensitive cells based on the gene expression pattern, AhR agonist and a compound exhibiting anti-tumor activity It makes it possible to predict sensitivity.

It is a figure which shows that the action spectrum in breast cancer of 150460 and 5F-203 corresponds. It is a figure which shows that CYP1A1 mRNA is induced | guided | derived in a 150460 sensitivity breast cancer, and is not induced | guided | derived in an insensitive breast cancer. It is a figure which shows that CYP1A1 protein is induced | guided | derived in a 150460 sensitivity breast cancer, and is not induced | guided | derived in an insensitive breast cancer. It is a figure which shows that the cell growth inhibitory effect of 150460 is canceled by knocking down AhR and CYP1A1 in 150460 sensitivity breast cancer SK-BR-3. The number of cells in each group after treatment with 0, 2, 10 μM 150460 and culturing for 48 hours was relative to the number of cells in the 150460 untreated group as 100. FIG. 150460 shows the growth inhibitory effect on the hepatoma cell line. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 30 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. 26 of the 266 genes selected in the present invention are shown. Whether sensitive cells or non-sensitive cells are highly expressed is also described. It is a figure which shows that the sensitive cell of 150460 and an insensitive cell can be clearly divided by the cluster analysis by 25 types of genes selected arbitrarily. It is a figure which shows that the nude rat subcutaneous transplant human breast cancer cell KPL-1 with unknown sensitivity is suggested to be a sensitive cell by the cluster analysis by 25 types of genes selected arbitrarily. It is a figure in which human breast cancer cell KPL-1 transplanted subcutaneously into nude rats is sensitive to 150460. The horizontal axis represents the number of days from the start date of administration, and the vertical axis represents the relative tumor volume. The control group is the symbol ◆, 150460-P 60 mg / kg 28 days continuous oral administration group is the symbol ■, 150460-P 30 mg / kg The 28 day continuous oral administration group is the symbol ▲, 150460-P 15 mg / kg 28 days continuous The oral administration group is indicated by the symbol ●.

As a result of DNA microarray analysis of more than 30,000 genes in various cells, there is a statistically significant difference in the expression level between cells that are AhR agonists and cells that are not sensitive to CYP1A1 activated compounds ( It was revealed that there are 266 genes having P <1.0 × 10 ⁻⁵ ). In the determination of sensitivity, the expression level of mRNA extracted from a cancer tissue isolated from a cancer patient is analyzed using means such as a gene chip. Next, based on the expression levels of 20 or more genes arbitrarily selected from the 266 genes, the sensitivity of cancer cells to a compound that is an AhR agonist and exhibits antitumor activity is determined.

  By measuring the expression level of any gene among these 266 genes, it is possible to determine the sensitivity to a compound that is an AhR agonist and is activated by CYP1A1. Furthermore, the sensitivity can be determined only by the expression levels of 20 or more genes arbitrarily selected from 266 genes. Details of the present invention will be described below.

  The 266 genes used in the present invention can be obtained by the following method. Here, the “expression amount” does not need to be an absolute amount but may be a relative amount. In addition, it is not always necessary to express numerically. For example, a case where a visual label such as a fluorescent label is used as a label to make a visual determination or the like corresponds to “measurement of expression level”.

-Production of DNA microarray-
A DNA microarray is prepared using the synthetic DNA. The DNA microarray is obtained by fixing a single-stranded oligo DNA consisting of a partial sequence of a target gene on a silicon substrate or a slide glass, and is sometimes called a DNA chip. It is used as a tool to comprehensively analyze the expression level of a gene complementary to a fixed single-stranded oligo DNA sequence, and it is possible to analyze 1 to several hundred thousand gene expression levels at once. It has become. The synthetic DNA here is a single-stranded oligo DNA complementary to a known gene, and is used for measuring the gene expression level in a cell. For example, synthetic DNA having a length of 25 to 100 bases is used, but is not necessarily limited thereto. As a method for producing a microarray, for example, synthetic DNA is spotted on a slide glass using an arrayer and then kept at 80 ° C. for 1 hour in a gas-phase incubator, and further using a UV crosslinker. Examples include, but are not necessarily limited to, a method of irradiating 120 mJ ultraviolet rays and immobilizing the spotted synthetic DNA on the surface of the slide glass.

-Extraction of total RNA-
Next, total RNA is extracted from the cancer cells. Extraction of total RNA can be performed according to the method described in (Chomczynski, P. and Mackey, K., BioTechniques 19 (6): 942-945 (1995)), but is not limited thereto. In the present invention, it is necessary to select one or more types of cells that are sensitive to a compound activated by CYP1A1 and one or more cells that are not sensitive as cancer cells from which total RNA is extracted. Examples of cancer cells that are sensitive to a compound activated by CYP1A1 that are AhR agonists include HCC1419, MDA-MB-175VII, MDA-MB-361, MDA-MB-453, MCF-7, R27. , ZR-75-1, CAMA1, SK-BR-3, ZR-75-30, T-47D and MDA-MB-468 and other breast cancer cells, and HepG2 and Hep3B hepatoma cells. Examples of cells that are not sensitive to a compound activated by CYP1A1 as AhR agonist include, for example, MDA-MB-157, MDA-MB-231, MDA-MB-134IV, UACC812, Hs0578T, MDA-MB Breast cancer cells such as -435s and HCC1937, liver cancer cells such as SK-Hep-1, HLF, HLE, PLC / PRF / 5 and Li7, and ovarian cancer cells such as AdrR. However, it is not limited to the cancer cells listed here.

-Purification of sample mRNA-
Specimen mRNA is purified from total RNA extracted from cancer cells. Although there are no limitations on the mRNA purification method and conditions, for example, the purification method described in (Goss TA, Bard M, Jarret HW., J. Chromatogr., 588 (1-2): 157-164 (1991)), etc. Is mentioned.

-Preparation of labeled cDNA-
Using the obtained mRNA for specimen as a template, a labeled cDNA can be prepared by using a nucleic acid with an arbitrary labeled molecule. As the labeling molecule, for example, cyanine 5-deoxyuridine phosphate (Cy5-dUTP), cyanine 3-deoxyuridine phosphate (Cy3-dUTP) and the like can be used in the present invention. Methods and conditions for producing labeled cDNA can be performed according to the method described in (Richter, A. et al., BioTechniques, 33 (3): 620-630 (2002)), but are not limited thereto. is not.

-Hybridization-
The obtained labeled cDNA is used as a detection probe to hybridize with the prepared DNA microarray. As a result, the labeled cDNA contained in the sample specifically hydrogen bonds with the complementary oligo DNA on the DNA microarray. Subsequently, other than labeled cDNA that specifically hybridized with the oligo DNA on the DNA microarray is removed by a washing operation. The conditions for the hybridization operation and the washing operation are appropriately determined. For example, the conditions described in (Schena, M. et al., Science, 270 (5235): 467-470 (1995)) are performed. Can do.

-Measurement of gene expression level-
After performing the hybridization operation and the washing operation, the expression level of each gene can be estimated by measuring the fluorescence intensity of the labeled cDNA bound to each synthetic DNA immobilized on the DNA microarray. Moreover, the relative gene expression level in the target cell can be calculated by using one or a plurality of arbitrary cells as control cells.

-Statistical processing-
The gene expression level of each gene is analyzed by statistical processing, and genes having a significant difference in expression level between the sensitive cell group and the insensitive cell group are extracted. As a statistical processing method, Student's t-test method is used, and a statistically significant method is used in the determination method of the present invention. Here, the statistical significance referred, between the sensitive group and insensitive group, and those wherein P <1.0 × 10 ^-5, in the present invention P <1.0 × 10 ^{- 7} is preferable, and P <1.0 × 10 ⁻⁹ is more preferable.

-Method for judging sensitivity-
The expression level of at least one of the genes in a cancer cell collected from a cancer patient by surgery or biopsy is measured. The number of genes to be measured may be any number as long as it is 1 or more, and any of the 266 genes may be used. Among these genes, it is preferable to use the expression level of any one or more of SEQ ID NOs: 1 to 30. Of these, it is more preferable to measure the expression levels of 20 or more genes.

  The expression level of each gene in the cell can be measured by measuring the mRNA level of each gene in the cell, and the mRNA level can be measured by a well-known method. For example, a DNA microarray in which an oligo DNA complementary to any gene used in the determination method of the present invention is immobilized is prepared. On the other hand, labeled cDNA is synthesized based on the mRNA of the specimen by the same method as the preparation of the labeled cDNA. The obtained labeled cDNA is reacted with the DNA microarray, and the labeled cDNA and the oligo DNA on the DNA microarray are hybridized. The expression level of the gene can be measured by washing after hybridization and measuring the labeling amount of each spot on the DNA microarray. The method for measuring the expression level of each gene in the specimen is not limited to this method, and any other method can be adopted as long as the expression level of the gene can be measured. For example, it is possible to measure each RNA in a specimen also by a real-time detection reverse transcription PCR method or a Northern blot method. A preferred embodiment includes a method in which labeled cDNA prepared from a specimen is hybridized with each oligo DNA immobilized on a microarray and the amount of the label is measured.

  Next, the expression level of each gene of the measured specimen is compared with the expression level of each gene in cells having and not sensitive to a compound activated by CYP1A1, which is an AhR agonist. The cells with and without sensitivity may be cultured cancer cell lines, or specimens from known responders and non-responders.

  Each specimen can be hierarchically clustered using the expression levels of these genes in cells with and without sensitivity as an index. Hierarchical cluster analysis can be performed using software such as “cluster” or “treeview”. Clusters constructed using gene expression levels with sufficiently high differences in expression levels between sensitive and insensitive groups can be clearly distinguished from sensitive and insensitive cell groups. Therefore, by performing a hierarchical cluster analysis with additional samples for which sensitivity is to be determined, it is possible to determine the drug sensitivity of the sample by determining whether the sample is distributed in a cluster of sensitive cells or a cluster of insensitive cells. Can be determined.

  According to the above method, whether the specimen is AhR agonist and sensitive to a compound exhibiting antitumor activity, that is, whether it is a responder or non-responder of treatment with a compound that is AhR agonist and exhibits antitumor activity. Can be determined.

  The compound whose sensitivity can be determined by the above-described method of the present invention is not limited as long as it is an AhR agonist and exhibits an antitumor effect, but specific examples include (5S, 7S) -7-methyl-3-. (3-Trifluoromethylphenyl) -5,6,7,8-tetrahydrocinnolin-5-ol (150460), 2- (4-amino-3-methylphenyl) -5-fluorobenzothiazole (5F-203) ) Or (5-amino-2,3-fluorophenyl) -6,8-difluoro-7-methyl-4H-1-benzopyran-4-one (NSC 686288). In the present invention, AhR agonist, compounds having the same mechanism of action and action as compounds activated by CYP1A1, CYP1A2, or CYP1B1, and derivatives thereof, or pharmacologically acceptable salts thereof Etc., the sensitivity can be determined and the therapeutic effect can be predicted by the determination method of the present invention. Here, the derivative is a compound in which one or a plurality of arbitrary substituents constituting a certain compound are substituted with arbitrary different substituents or atoms, and exhibits a medicinal effect of similar properties. Here, the number of substituents is not particularly limited, but is preferably 5 or less, and examples of the substituent include lower alkyl groups having 1 to 6 carbon atoms, halogens, amino groups, hydroxyl groups, nitro groups, carboxyl groups, etc. Although the lower alkyl group of Formula 1-6 can be mentioned, it is not limited to these. Examples of pharmacologically acceptable salts include, but are not limited to, mesylate, hydrochloride, sulfate, nitrate, and the like.

  The microarray of the present invention refers to a DNA microarray in which one or more oligo DNAs complementary to the full length of a gene or a partial sequence thereof used in the determination method of the present invention are immobilized. Among the above genes, those obtained by immobilizing any one or more of SEQ ID NOS: 1 to 30 or oligo DNA complementary to a partial sequence thereof are preferred, and those comprising 20 or more species are more preferred. As used herein, complementary oligo DNA having a length of 25 to 100 bases is generally used, but is not necessarily limited thereto.

  The kit of the present invention is an AhR agonist of the present invention and includes a microarray for determining the sensitivity of cancer cells to a compound exhibiting antitumor activity, a derivative thereof, or a pharmacologically acceptable salt thereof. , Reagents for hybridization, reaction solutions, washing solutions, and the like.

  The primer set of the present invention refers to the AhR agonist of the present invention, a gene used for determining the sensitivity of cancer cells to a compound exhibiting antitumor activity, a derivative thereof, or a pharmacologically acceptable salt thereof by PCR. Oligo DNA for amplification by In general, oligo DNA having a length of 17 to 40 bases is used, but is not necessarily limited thereto. These oligo DNAs can be synthesized by a known method such as a phosphoramidite method. The amount of the target gene can be quantified by real-time PCR using primers using cDNA synthesized by reverse transcription reaction from mRNA derived from the specimen as a template. When the real-time PCR method is performed, a DNA intercalator fluorescent dye such as Cyber Green is used to measure the amount of PCR product produced, and MX3000P (manufactured by Stratagene) can be used as a measuring device. However, these conditions can be appropriately changed as long as the experiment can be reproduced. Cases where the invention is carried out under the changed conditions are also included in the scope of the present invention. Since the target gene can be quantified by this method, sensitivity can be determined according to the method described above.

  The antibody set of the present invention refers to the AhR agonist of the present invention, which immunizes a gene product used for determining the sensitivity of cancer cells to a compound exhibiting antitumor activity, a derivative thereof, or a pharmacologically acceptable salt thereof. Refers to the antibody used to stain. The antibody may be anything, such as a monoclonal antibody or a polyclonal antibody, as long as it can specifically recognize the antigen. These antibodies can also be obtained by immunizing mice, rabbits, goats and the like with antigens. This antigen may be the full length of the target gene product or a part thereof. By immunostaining the tumor tissue with these antibodies, it becomes possible to know the expression level of the target gene, so that sensitivity can be discriminated in accordance with the method described below.

  Hereinafter, the present invention will be described more specifically based on examples. However, the present invention is not limited to the following examples.

Example 1 Demonstration that 150460 AhR agonist action is required for anti-tumor effect

1. Induction of CYP1A1 mRNA by 150460 150460 sensitive breast cancer cell lines MCF-7, T-47D, SK-BR-3, MDA-MB-453, and 150460 insensitive breast cancer cell lines MDA-MB-435S, Hs0578T, MDA -MB-231 was seeded in a culture dish having a diameter of 90 mm each with 1 million MB-231, cultured overnight in RPMI 1640 medium containing 10% FBS, treated with 150460 at a final concentration of 0 and 2.5 μM, and cultured for 3 hours. . Subsequently, after removing the culture solution, the cells were washed with Phosphate Buffered Saline (PBS), and 1 mL of ISOGEN solution (manufactured by Nippon Gene) was added to lyse the cells. 250 μL of chloroform was added to the lysed cells, and the mixture was vigorously stirred and centrifuged at 15,000 rpm for 3 minutes at 4 ° C. to remove proteins. 500 μL of the supernatant was collected, an equal amount of isopropanol was added, and the mixture was incubated at room temperature for 10 minutes, and then centrifuged at 12,000 rpm for 10 minutes at 4 ° C. to precipitate RNA. The precipitated RNA was washed with 1 mL of 75% ethanol, ethanol was removed, and water was added to obtain an RNA solution. The RNA solution was assayed for concentration and purity using absorbance.

  CDNA synthesis was performed using 1 μg of RNA each. Specifically, 1 μg of RNA and 2 μL of 50 μM oligo (dT) 20 solution (Invitrogen) were incubated at 65 ° C. for 5 minutes, and then 1 μL of Superscript III reverse transcriptase (Invitrogen), 4 μL of 5 × concentration buffer (SuperScript III) (Attached to reverse transcriptase) 1 μL RNase inhibitor (Invitrogen), 1 μL 0.1 M dithiothreitol, 4 μL 2.5 mM dNTP, and water to make a total volume of 20 μL, gently After pipetting, the mixture was kept at 50 ° C. for 50 minutes to synthesize cDNA.

  Subsequently, the expression level of CYP1A1 gene contained in the cDNA was quantified. To 1 μL of the obtained cDNA, 15 μL of TaqMan Universal PCR Master Mix (Applied Biosystems), 1.5 μL of CYP1A1 quantification TaqMan probe (Applied Biosystems), 12.5 μL of water were added and gently stirred. Thereafter, the gene was amplified by performing 40 cycles of 95 ° C. for 15 seconds and 60 ° C. for 1 minute using a real-time PCR apparatus MX3000P (manufactured by Stratagene). For each cell, the point where the gene was logarithmically amplified was selected and the Ct value was determined. As a control gene, a glyceraldehyde triphosphate dehydrogenase (hereinafter referred to as GAPDH) gene was used, and the Ct value was determined by the same method. For the 150460-treated group and the non-treated group of each cell, the value obtained by subtracting the CPD value of GAPDH from the Ct value of the CYP1A1 gene was obtained as a ΔCt value and compared with each other, thereby comparing the relative CYP1A1 in each group. The expression level was estimated. FIG. 2 shows the expression level of the 150460 treatment group when the expression level of the CYP1A1 gene in the 150460 non-treatment group is defined as 100. As a result, it was revealed that induction of CYP1A1 gene expression was observed in 150460-sensitive cancer but not in 150460-insensitive cancer.

2. Induction of CYP1A1 protein by 150460 Subsequently, whether CYP1A1 protein was induced by 150460 was examined. 150460, sensitive breast cancer cell lines MCF-7, T-47D, SK-BR-3, MDA-MB-453, and 150460 insensitive breast cancer cell lines MDA-MB-435S, Hs0578T, MDA-MB-231, respectively. One million pieces were seeded in a culture dish having a diameter of 90 mm and cultured overnight in RPMI1640 medium containing 10% FBS, and then treated with 150460 at a final concentration of 0 and 1 μM and cultured for 6 hours. After removing the medium, the cells were washed with 10 mL of PBS, and the cells were collected with a scraper. To each cell, 200 μL of cell lysate (50 mM Tris-HCl (pH 7.5), 300 mM sodium chloride, 0.5% Triton X-100) was added, stirred by pipetting, and incubated on ice for 30 minutes. did. Subsequently, the mixture was centrifuged at 15000 rpm for 30 minutes at 4 ° C., and the supernatant was recovered as a protein fraction. The protein fraction was subjected to protein quantification using a BCA protein assay kit (Pierce) to determine the concentration. 20 μg of each protein was fractionated by 10% SDS polyacrylamide gel electrophoresis, and transferred to a PVDF membrane using a blotting apparatus. After blocking for 1 hour at room temperature with a TBS-T solution (20 mM Tris-HCl (pH 7.5), 150 mM sodium chloride, 0.05% Tween 20) containing 5% skim milk, 0.2 μg / mL anti-CYP1A1 antibody (Sc-20772, manufactured by Santa Cruz) was added and the reaction was carried out at 4 ° C. overnight. After washing 3 times with TBS-T, an HRP-conjugated anti-rabbit IgG antibody (manufactured by GE Healthcare) was added as a secondary antibody and reacted at room temperature for 1 hour. After washing three times with TBS-T, chemiluminescence was performed using an ECL western blotting detection system (manufactured by GE Healthcare) to expose the X-ray film. The results of Western blotting are shown in FIG. From this result, it was clarified that induction of CYP1A1 protein occurred in 150460-sensitive cells and not in insensitive cells.

3. Proof that AhR and CYP1A1 are required for the anti-tumor effect of 150460 Subsequently, whether CYP1A1 and AhR were required for the anti-tumor effect of 150460 was examined. 150,000-sensitive breast cancer cell line SK-BR-3 was seeded on a 24-well culture plate at a rate of 50,000 cells and cultured overnight in RPMI1640 medium containing 10% FBS, and then AhRsiRNA (Invitrogen), CYP1A1 siRNA (Invitrogen) ) As a negative control, non-silencing siRNA (manufactured by Invitrogen) was introduced by the following method. Three types of AhRsiRNA and CYP1A1 siRNA were used. That is, 20 pmol of siRNA was mixed with 1 μL of Lipofectamine 2000 (manufactured by Invitrogen) and 100 μL of OPTI-MEM (manufactured by Invitrogen), incubated at room temperature for 20 minutes, and then administered to each culture well. After administration, the cells were cultured for 24 hours, and after knocking down each gene, 150460 at a final concentration of 0, 2, 10 μM was treated, and further cultured for 48 hours. Subsequently, the culture broth was extracted, fixed with methanol, and then stained with methylene blue to measure the number of cells. It was examined whether the cell growth inhibitory effect by 150460 and each siRNA could cancel the action. The result is shown in FIG. Since both AhR and CYP1A1 siRNA strongly suppressed the cell growth inhibitory effect of 150460, it was revealed that AhR and its downstream CYP1A1 are necessary for the cell growth inhibitory action of 150460.

Example 2 Anti-tumor effect of 150460 150460 shows a cell growth inhibitory action on a plurality of breast cancer cell lines, and whether or not such an action was observed in other cell types was examined using a liver cancer cell line. Liver cancer cell lines HLE, HLF, HuH6, HuH7, PLC / PRF / 5, SK-Hep1, HT17, Hep3B, and HepG2 strains were seeded in a 96-well culture dish and cultured overnight in RPMI1640 medium containing 10% FBS. Subsequently, 150460 having a final concentration of 0, 0.016, 0.08, 0.4, 2, 10 μM was treated and further cultured for 72 hours. Subsequently, the culture solution was extracted, fixed with methanol, and then stained with methylene blue to measure the number of cells, and the cell growth inhibitory effect of 150460 was examined. The result is shown in FIG. It was revealed that the HuH6, HT17, Hep3B, and HepG2 strains were sensitive to 150460, and that 150460 had a growth inhibitory effect in cell lines other than breast cancer.

Example 3 Production of Microarray A human gene fragment library (Microdiagnostic) was placed on a slide glass (Matsunami Glass Kogyo, S9115) using an ultra-small volume dispensing device (MicroDiagnostic, GeneMatrixMaker-R). To make a microarray. The microarray slide glass was inserted into a slide staining basket (manufactured by Matsunami Glass Co., Ltd., B-60) and kept warm in a gas-phase incubator (manufactured by Stabil Life Science Co., Hybridization Oven) at 80 ° C. for 1 hour. Further, 120 mJ of ultraviolet light was irradiated using a UV crosslinker (Hoefer, UVC500), and the spotted synthetic DNA was immobilized on the surface of the slide glass.

1. Selection of genes to be used for determination-Preparation of total RNA-
For analysis, among the 150460 sensitive cancer cells, MCF-7, MDA-MB-361, SK-BR-3, T-47D, MDA-MB-453, MDA-MB-175-VII, ZR-75- 1, R27, HCC1419, ZR-75-30, and 11 types of CAMA1 were used. Among 150460 insensitive cells, MDA-MB-435S, MDA-MB-134IV, Hs0578T, MDA-MB-231, MDA-MB-157 and UACC812 are used as breast cancer, and HLE, SK- 12 types of AdrR were used as Hep-1, PLC / PRF / 5, HLF and Li7, and ovarian cancer. These cells were cultured in an RPMI1640 medium containing 10% FBS in a culture flask having a bottom area of 175 cm ² , and when the cell density reached 80-90%, the medium was quickly removed and 10.5 mL After lysing cells by adding ISOGENreagent (manufactured by Nippon Gene), total RNA was extracted according to the attached manual. The purity of 100 g or more of total RNA obtained according to the protocol was confirmed by measuring absorbance at 260 nm and 280 nm with a spectrophotometer. Furthermore, after confirming the presence or absence of RNA degradation by denaturing agarose gel electrophoresis, the inventors proceeded to the following experiment.

-Synthesis of labeled cDNA-
Sample mRNA was obtained by extracting polyadenylic acid [poly (A)] + RNA from total RNA extracted from each cell using a Poly (A) Pure kit (Ambion). 2.0 μg of each mRNA was collected and labeled and hybridized kit (manufactured by Microdiagnostic), reverse transcriptase SuperScript II (manufactured by Invitrogen), and cyanine 5-deoxyuridinetriphosphate (Cyanine5-dUTP) (manufactured by PerkinElmer) ) To prepare labeled cDNA for each cell specimen. As a control, cDNA was prepared by labeling a mixed sample of equal amounts of mRNA of all cells used with Cyanine 3-deoxyuridine triphosphate (Cyanine 3-dUTP) (PerkinElmer).

-Preparation of labeled probe-
These labeled cDNAs, that is, each cell specimen labeled with Cyanine5 and a control labeled with Cyanine3 are mixed in the same test tube, purified by Microcon YM30 (Millipore), and finally labeled with nucleic acid / hybridization. It was prepared to 15 μL using a hybridization buffer and pure water attached to the reagent (manufactured by Microdiagnostic).

-Hybridization-
This solution was heat denatured by heating at 99 ° C. for 5 minutes, and allowed to stand at room temperature for 5 minutes. Then, the solution was dropped on the DNA spotted area of the microarray slide glass, and a cover glass of 24 mm × 40 mm (Matsunami Glass Industrial Co., Ltd.) Made of NEO cover glass) and stored in a hybridization cassette (manufactured by Micro Diagnostics). The hybridization cassette storing the microarray was placed in a gas-phase thermostat (manufactured by Sanyo Biomedica Co., Ltd., MIR262) and kept warm at 42 ° C. for 20 hours.

-Washing-
The hybridization cassette was taken out of the vapor phase incubator, and the slide glass was washed using a hybridization washing solution attached to a nucleic acid labeling / hybridization reagent (manufactured by Microdiagnostic) according to the protocol recommended by the company.

-Detection and quantification of hybridization-
The microarray slide glass thus hybridized and washed was simultaneously measured for fluorescence signals at wavelengths of 635 nm and 532 nm using a scanner (Axon Instrument, GenePix 4000B). Optical evaluation was performed using analysis software (Axon Instrument, GenePix Pro) attached to the scanner, and the relative expression level relative to the control was calculated from the Cyanine3: Cyanine5 ratio for each sample.

-Statistical processing-
Next, in order to select genes useful for discriminating and separating sensitive cells from insensitive cells, the probability of Student's t-distribution is expressed with respect to the logarithmically converted relative expression level of each gene between sensitive cells and non-sensitive cells. The returned TDIST function was implemented to calculate the P value.

As a result, 266 genes having permutation P values of less than 1.0 × 10 ⁻⁵ were extracted as candidates (FIGS. 6 to 14). Of these 266 genes, 144 genes had increased expression levels in insensitive cells compared to sensitive cells, and 122 genes had decreased expression levels.

2. Clustering by Genes with Large Significant Differences Hierarchical cluster analysis was performed using 25 genes of SEQ ID NOs: 1-25. Hierarchical cluster analysis is Software developed by Eisen and available from the web on the Internet, “cluster” and “treeview” (http: //genome-www5/stanford.edu/MicroArray/SMD/retech.html). Prior to applying the clustering algorithm, the fluorescence ratio for each spot was log transformed, and then the data for each sample was median centered to remove experimental bias. As a result, the cells were clearly separated into sensitive and insensitive cells by analysis using these gene sets (FIG. 15).

Example 4 Prediction of 150460 Sensitivity of Tumor with Unknown 150460 Sensitivity Total RNA was obtained from breast cancer cell line KPL-1 transplanted into nude rats for the purpose of examining whether the sensitivity of tumors with unknown 150460 sensitivity can be determined. Microarray profiling was performed. By adding KPL-1 to the cluster analysis made in FIG. 15, it was predicted that KPL-1 was sensitive to 150460 (FIG. 16). Subsequently, 150460-P was administered to KPL-1 cells transplanted into nude rats, and 150460 sensitivity was confirmed (FIG. 17). From this, it became clear that by using this sensitivity determination method, it is possible to determine the sensitivity in advance even for a tumor whose sensitivity is unknown. Specific experimental examples are shown below.

1. Preparation and microarray analysis of total RNA from in vivo tumors Human breast cancer strain KPL-1 with unknown sensitivity of 150460 was obtained from nude mice (BALB / cAJcl-nu / nu female; manufactured by CLEA Japan, Inc.) at our facility. Transplanted and maintained subcutaneously on the side. Then, the KPL-1 maintained for subculture was transplanted subcutaneously into nude rats (F344 / NJcl-rnu / run female; manufactured by CLEA Japan, Inc.) and the proliferated tumor was extracted and frozen in liquid nitrogen in advance. It was put in a 15 mL tube to which ISOGEN reagent (manufactured by Nippon Gene) was added. The frozen tumor placed in the tube was thoroughly crushed using Polytron (manufactured by KINEMATICA), and then total RNA was extracted according to the protocol. The total RNA obtained was checked for purity and concentration by measuring absorbance at 260 nm and 280 nm with a spectrophotometer. Furthermore, after confirming the presence or absence of RNA degradation by denaturing agarose gel electrophoresis, the inventors proceeded to the following experiment.

  In the same manner as in Experimental Example 2 of Example 3, gene expression analysis by microarray was performed on KPL-1 implanted subcutaneously in nude rats. Of the data obtained there, 25 gene expression levels of SEQ ID NOs: 1 to 25 were selected. The subjects to be analyzed include MCF-7, MDA-MB-361, SK-BR-3, T-47D, MDA-MB-453, MDA-MB-175-VII, ZR-75 as sensitive breast cancer cells. -11, R27, HCC1419, ZR-75-30, and CAMA1 were used. Among 150460 insensitive cells, MDA-MB-435S, MDA-MB-134IV, Hs0578T, MDA-MB-231, MDA-MB-157 and UACC812 are used as breast cancer, and HLE, SK- 12 types of AdrR were used as Hep-1, PLC / PRF / 5, HLF and Li7, and ovarian cancer. Based on the data of 11 types of sensitive cells and 12 types of insensitive cells regarding the expression levels of these 25 selected genes, hierarchical cluster analysis was performed for KPL-1. Hierarchical cluster analysis was performed using “cluster” and “treeview” as in Experimental Example 2. Prior to applying the clustering algorithm, the fluorescence ratio for each spot was log transformed, and then the data for each sample was median centered to remove experimental bias. As a result, analysis using these gene sets suggested that KPL-1 is a sensitive tumor (FIG. 16).

2. Evaluation of 150460 sensitivity of in vivo tumors To determine the sensitivity of KPL-1 to 150460, an antitumor test was conducted against KPL-1 implanted subcutaneously in nude rats. Nude rats transplanted with KPL-1 were randomly assigned to each group based on tumor volume and body weight on the day of administration. The tumor volume (TV (mm ³ )) was calculated from the following formula using a digital caliper, using a major axis (Lmm) and a minor axis (Wmm).

  Tumor volume is calculated, defective ones are excluded from the tumor shape and engraftment position, individuals are removed by multivariable using tumor volume and body weight, and then block assignment by multivariable (SAS system preclinical) Package Ver.5.0 (manufactured by SAS Institute Japan Co., Ltd.) was performed, and 6 groups were assigned to 4 groups. The 150460-P non-administered group and 150460-P were divided into groups in which 60 mg / kg, 30 mg / kg and 15 mg / kg were orally administered once a day for 28 days, respectively. A tumor growth curve over time was prepared from the relative tumor volume with the tumor volume on the day of administration as 1. As a result, KPL-1 was shown to be sensitive to 150460 (FIG. 17). From this, it was shown that 150460 sensitivity of cells with unknown sensitivity can be determined by sensitivity determination using the above gene set.

  Using the determination method and kit of the present invention, mRNA extracted from cancer cells isolated from cancer patients by means of surgery or biopsy, etc. is used as a sample, and its gene expression pattern is analyzed using a gene chip or the like to detect sensitive cells and non-sensitive cells. By comparing gene expression patterns with sensitive cells, the sensitivity to drugs can be predicted.

SEQ ID NO: 1: SPDEF cDNA SEQ ID NO: 2: MSN cDNA sequence SEQ ID NO: 3: PROM2 cDNA sequence SEQ ID NO: 4: FLJ33235 cDNA sequence SEQ ID NO: 5: RAB25 cDNA sequence SEQ ID NO: 6: STARD10 cDNA sequence number 7: cDNA sequence of XBP1 SEQ ID NO: 8: cDNA sequence of CREB3L4 SEQ ID NO: 9: cDNA sequence of IGF2BP2 SEQ ID NO: 10: cDNA sequence of FLJ10444 SEQ ID NO: 11: cDNA sequence of CAV2 SEQ ID NO: 12: cDNA sequence of FLJ35418 ANXA1 cDNA sequence SEQ ID NO: 14: TOB1 cDNA sequence SEQ ID NO: 15: GRHL2 cDNA sequence SEQ ID NO: 16: DDR1 cDNA sequence SEQ ID NO: 17: FLJ41692 cDNA sequence SEQ ID NO: 18 YDE1 cDNA SEQ ID NO: 19: PRSS8 cDNA sequence SEQ ID NO: 20: CLDN3 cDNA sequence SEQ ID NO: 21: CMTM3 cDNA sequence SEQ ID NO: 22: POPDC3 cDNA sequence SEQ ID NO: 23: EMP3 cDNA sequence SEQ ID NO: 24: KIAA0182 cDNA sequence SEQ ID NO: 25: TRPS1 cDNA sequence SEQ ID NO: 26: CCDC82 cDNA sequence SEQ ID NO: 27: STK17A cDNA sequence SEQ ID NO: 28: RBM35A cDNA sequence SEQ ID NO: 29: C1orf172 cDNA sequence SEQ ID NO: 30: JUP cDNA sequence SEQ ID NO: 31: cDNA sequence of FLJ10960 SEQ ID NO: 32: cDNA sequence of PSD4 SEQ ID NO: 33: cDNA sequence of ARRDC1 SEQ ID NO: 34: cDNA sequence of RGL2 SEQ ID NO: 35: L X1L cDNA SEQ ID NO: 36: SPINT1 cDNA SEQ ID NO: 37: CAV1 cDNA SEQ ID NO: 38: LUZP1 cDNA SEQ ID NO: 39: IRX5 cDNA SEQ ID NO: 40: SH3KBP1 cDNA SEQ ID NO: 41: LEPRE1 cDNA sequence SEQ ID NO: 42: WBSCR21 cDNA sequence SEQ ID NO: 43: ARGGAP8 cDNA sequence SEQ ID NO: 44: EMP2 cDNA sequence SEQ ID NO: 45: MAL2 cDNA sequence SEQ ID NO: 46: BMP1 cDNA sequence SEQ ID NO: 47: GALNT6 cDNA sequence SEQ ID NO: 48: cDNA sequence of PRKCA SEQ ID NO: 49: cDNA sequence of CA12 SEQ ID NO: 50: cDNA sequence of EFNA1 SEQ ID NO: 51: cDNA sequence of CLDN7 SEQ ID NO: 52: cDN of ACSL4 A sequence SEQ ID NO: 53: cDNA sequence of RHOB SEQ ID NO: 54: cDNA sequence of CDC42SE1 SEQ ID NO: 55: cDNA sequence of ST3GAL2 SEQ ID NO: 56: cDNA sequence of LARP6 SEQ ID NO: 57: cDNA sequence of PSMB9 SEQ ID NO: 58: cDNA sequence of SLC37A1 SEQ ID NO: 59: cDNA sequence of CARD10 SEQ ID NO: 60: cDNA sequence of TRIM5 SEQ ID NO: 61: cDNA sequence of SERENBP1 SEQ ID NO: 62: cDNA sequence of FADS3 SEQ ID NO: 63: cDNA sequence of GATA3 SEQ ID NO: 64: cDNA sequence of C10orf57 65: cDNA sequence of ANTXR1 SEQ ID NO: 66: cDNA sequence of ENSA SEQ ID NO: 67: cDNA sequence of FLJ33850 SEQ ID NO: 68: cDNA sequence of F11R SEQ ID NO: 69: HEG1 cDNA sequence SEQ ID NO: 70: cDNA sequence of SMAD3 SEQ ID NO: 71: cDNA sequence of KRT10 SEQ ID NO: 72: cDNA sequence of FLJ31874 SEQ ID NO: 73: cDNA sequence of TACSTD2 SEQ ID NO: 74: cDNA sequence of ZNF217 SEQ ID NO: 75: cDNA sequence of FLJ43436 SEQ ID NO: 76: cDNA sequence of PBX1 SEQ ID NO: 77: cDNA sequence of FLJ20653 SEQ ID NO: 78: cDNA sequence of BSPRY SEQ ID NO: 79: cDNA sequence of DNAPTP6 SEQ ID NO: 80: cDNA sequence of DENND2D SEQ ID NO: 81: cDNA sequence number of KIAA1543 82: cDNA sequence of FLJ22945 SEQ ID NO: 83: cDNA sequence of GPR160 SEQ ID NO: 84: cDNA sequence of LOXL2 SEQ ID NO: 85: cDNA sequence of LYN No. 86: KIAA0802 cDNA SEQ ID NO: 87: PHLDB2 cDNA SEQ ID NO: 88: SMARCD2 cDNA SEQ ID NO: 89: BIN1 cDNA SEQ ID NO: 90: NBEAL2 cDNA SEQ ID NO: 91: MAPK12 cDNA SEQ ID NO: 92 : AGPS cDNA SEQ ID NO: 93: FLJ39770 cDNA sequence SEQ ID NO: 94: ICA1 cDNA sequence SEQ ID NO: 95: DCUN1D2 cDNA sequence SEQ ID NO: 96: KIF3C cDNA sequence SEQ ID NO: 97: GPD1L cDNA sequence SEQ ID NO: 98: FLJ30550 SEQ ID NO: 98: KIAA1324 cDNA sequence SEQ ID NO: 100: FLJ12354 cDNA sequence SEQ ID NO: 101: TSPAN15 cDNA sequence SEQ ID NO: 102: C UL4B cDNA SEQ ID NO: 103: TAX1BP3 cDNA SEQ ID NO: 104: FLJ30842 cDNA SEQ ID NO: 105: ACTN1 cDNA SEQ ID NO: 106: ABCC4 cDNA SEQ ID NO: 107: TRIM3 cDNA SEQ ID NO: 108: FLJ39084 cDNA sequence SEQ ID NO: 108: MAP7 cDNA sequence SEQ ID NO: 110: OSMR cDNA sequence SEQ ID NO: 111: FXYD3 cDNA sequence SEQ ID NO: 112: COTL1 cDNA sequence SEQ ID NO: 113: KRT19 cDNA sequence SEQ ID NO: 114: EPN3 cDNA sequence SEQ ID NO: 115: cDNA sequence of IGSF9 SEQ ID NO: 116: cDNA sequence of EFHD1 SEQ ID NO: 117: cDNA sequence of CLN3 SEQ ID NO: 118: c of FLJ30953 NA sequence SEQ ID NO: 119: SEMA3F cDNA sequence SEQ ID NO: 120: INPP1 cDNA sequence SEQ ID NO: 121: DCBLD2 cDNA sequence SEQ ID NO: 122: DHRS13 cDNA sequence SEQ ID NO: 123: VIM cDNA sequence SEQ ID NO: 124: SIGIRR cDNA sequence SEQ ID NO: 125: cDNA sequence of IRF1 SEQ ID NO: 126: cDNA sequence of TRIO SEQ ID NO: 127: cDNA sequence of TMEM132A SEQ ID NO: 128: cDNA sequence of FLJ20913 SEQ ID NO: 129: cDNA sequence of SCYL3 SEQ ID NO: 130: cDNA sequence number of SYNGR2 131: cDNA sequence of ITGA5 SEQ ID NO: 132: cDNA sequence of SH3YL1 SEQ ID NO: 133: cDNA sequence of WWTR1 SEQ ID NO: 134: cDNA sequence of QKI No. 135: cDNA sequence of FLJ21563 SEQ ID NO: 136: cDNA sequence of IGFBP5 SEQ ID NO: 137: cDNA sequence of FN1 SEQ ID NO: 138: cDNA sequence of FLJ35677 SEQ ID NO: 139: cDNA sequence of LOC387882 SEQ ID NO: 140: cDNA sequence of APBB1 SEQ ID NO: 141 : CDNA sequence of FLJ30869 SEQ ID NO: 142: cDNA sequence of TAP2 SEQ ID NO: 143: cDNA sequence of BAG2 SEQ ID NO: 144: cDNA sequence of FLJ11232 SEQ ID NO: 145: cDNA sequence of EXT1 SEQ ID NO: 146: cDNA sequence of PPA2 SEQ ID NO: 147: HSD17B11 CDNA sequence SEQ ID NO: 148: MYL9 cDNA sequence SEQ ID NO: 149: MAP4K4 cDNA sequence SEQ ID NO: 150: ABCA2 cDNA sequence sequence No. 151: TFPI cDNA sequence SEQ ID NO: 152: INADL cDNA sequence SEQ ID NO: 153: LYPD3 cDNA sequence SEQ ID NO: 154: GLS cDNA sequence SEQ ID NO: 155: PATZ1 cDNA sequence SEQ ID NO: 156: FLJ21175 cDNA sequence SEQ ID NO: 157 : CDNA sequence of FLJ10500 SEQ ID NO: 158: cDNA sequence of C14orf147 SEQ ID NO: 159: cDNA sequence of CORO1C SEQ ID NO: 160: cDNA sequence of FLJ22131 SEQ ID NO: 161: cDNA sequence of NPDC1 SEQ ID NO: 162: cDNA sequence of FOXO3 SEQ ID NO: 163: ALDH3B2 CDNA sequence SEQ ID NO: 164: COMT cDNA sequence SEQ ID NO: 165: EPCAM cDNA sequence SEQ ID NO: 166: CFL2 cDNA sequence SEQ ID NO: 67: cDNA sequence of ITGB1 SEQ ID NO: 168: cDNA sequence of COL4A1 SEQ ID NO: 169: cDNA sequence of DCXR SEQ ID NO: 170: cDNA sequence of ST14 SEQ ID NO: 171: cDNA sequence of ANXA5 SEQ ID NO: 172: cDNA sequence of IGF2BP3 SEQ ID NO: 173: PDCD4 cDNA sequence SEQ ID NO: 174: RELB cDNA sequence SEQ ID NO: 175: MCFD2 cDNA sequence SEQ ID NO: 176: FMNL2 cDNA sequence SEQ ID NO: 177: TYMS cDNA sequence SEQ ID NO: 178: FOXA1 cDNA sequence SEQ ID NO: 179: PDLIM7 cDNA sequence SEQ ID NO: 180: AKAP12 cDNA sequence SEQ ID NO: 181: IGFBP2 cDNA sequence SEQ ID NO: 182: KRT17 cDNA sequence SEQ ID NO: 183: TMC4 cD A sequence SEQ ID NO: 184: cDNA sequence of ART4 SEQ ID NO: 185: cDNA sequence of TMEM183A SEQ ID NO: 186: cDNA sequence of PIK3CD SEQ ID NO: 187: cDNA sequence of OPTN SEQ ID NO: 188: cDNA sequence of JTB SEQ ID NO: 189: cDNA sequence of DZIP1 SEQ ID NO: 190: MET cDNA SEQ ID NO: 191: FLJ23431 cDNA sequence SEQ ID NO: 192: BTN3A3 cDNA sequence SEQ ID NO: 193: FLJ11628 cDNA sequence SEQ ID NO: 194: SPTBN2 cDNA sequence SEQ ID NO: 195: EPHA1 cDNA sequence SEQ ID NO: 196: cDNA sequence of FLJ33892 SEQ ID NO: 197: COL6A2 cDNA sequence SEQ ID NO: 198: TLN1 cDNA sequence SEQ ID NO: 199: IFI44 cDNA sequence sequence Column number 200: cDNA sequence of MAP7D1 SEQ ID NO: 201: cDNA sequence of SOAT1 SEQ ID NO: 202: cDNA sequence of FLJ32204 SEQ ID NO: 203: cDNA sequence of IGFBP7 SEQ ID NO: 204: cDNA sequence of CNTNAP1 SEQ ID NO: 205: cDNA sequence of C9orf30 206: cDNA sequence of SLC44A2 SEQ ID NO: 207: cDNA sequence of HADH SEQ ID NO: 208: cDNA sequence of ZNF385A SEQ ID NO: 209: cDNA sequence of FLJ32108 SEQ ID NO: 210: cDNA sequence of FLJ11417 SEQ ID NO: 211: cDNA sequence of TIMP1 SEQ ID NO: 212: CDNA sequence of NAV2 SEQ ID NO: 213: cDNA sequence of TAP1 SEQ ID NO: 214: cDNA sequence of ECHDC1 SEQ ID NO: 215: cDN of RAB3D SEQ ID NO: 216: cDNA sequence of FAM126A SEQ ID NO: 217: cDNA sequence of KIAA0754 SEQ ID NO: 218: cDNA sequence of COL6A1 SEQ ID NO: 219: cDNA sequence of CTNND1 SEQ ID NO: 220: cDNA sequence of ARGGAP23 SEQ ID NO: 221: cDNA sequence of AURKB No. 222: LOC151361 cDNA sequence SEQ ID NO: 223: TIMP2 cDNA sequence SEQ ID NO: 224: FAM57A cDNA sequence SEQ ID NO: 225: SPRY2 cDNA sequence SEQ ID NO: 226: CD44 cDNA sequence SEQ ID NO: 227: TGFBI cDNA sequence SEQ ID NO: 228 : CDNA sequence of FSCN1 SEQ ID NO: 229: cDNA sequence of PIK3R3 SEQ ID NO: 220: cDNA sequence of DHTKD1 SEQ ID NO: 231: C10orf 16 cDNA sequence SEQ ID NO: 232: CCDC50 cDNA sequence SEQ ID NO: 233: MGC16291 cDNA sequence SEQ ID NO: 234: TRIP10 cDNA sequence SEQ ID NO: 235: MACF1 cDNA sequence SEQ ID NO: 236: AKT3 cDNA sequence SEQ ID NO: 237: KIRREL cDNA sequence SEQ ID NO: 238: VCL cDNA sequence SEQ ID NO: 239: F2R cDNA sequence SEQ ID NO: 240: SCHIP1 cDNA sequence SEQ ID NO: 241: ADM cDNA sequence SEQ ID NO: 242: EPS8L1 cDNA sequence SEQ ID NO: 243: ADAM15 cDNA sequence SEQ ID NO: 244: cDNA sequence of FLJ10768 SEQ ID NO: 245: cDNA sequence of NRP1 SEQ ID NO: 246: cDNA sequence of FLJ31090 SEQ ID NO: 247: cDN of ARL4C A sequence SEQ ID NO: 248: cDNA sequence of FLJ90460 SEQ ID NO: 249: cDNA sequence of CCDC88A SEQ ID NO: 250: cDNA sequence of SLC41A3 SEQ ID NO: 251: cDNA sequence of NFE2L3 SEQ ID NO: 252: cDNA sequence of FLJ14044 SEQ ID NO: 253: cDNA sequence of SGK1 SEQ ID NO: 254: ABCG1 cDNA SEQ ID NO: 255: PARVA cDNA sequence SEQ ID NO: 256: FLJ45204 cDNA sequence SEQ ID NO: 257: FLJ34580 cDNA sequence SEQ ID NO: 258: GLT25D1 cDNA sequence SEQ ID NO: 259: UBE2E3c DNA sequence SEQ ID NO: 260: SLC25A1 cDNA sequence number 261: FLJ41966 cDNA sequence number 262: FLJ32235 cDNA sequence number 263: PPP2R5A cDNA sequence SEQ ID NO 264: HIG2 cDNA sequence SEQ ID NO 265: RNF208 cDNA sequence SEQ ID NO 266: TMEM141 cDNA sequences

Claims (8)

A compound, derivative thereof, which is an AhR agonist and has antitumor activity, comprising a step of measuring the expression level of one or more genes selected from a gene group consisting of 266 genes represented by SEQ ID NOs: 1 to 266, or A method for determining the sensitivity of a cancer cell to a pharmacologically acceptable salt thereof. The compound is (5S, 7S) -7-methyl-3- (3-trifluoromethylphenyl) -5,6,7,8-tetrahydrocinnolin-5-ol, 2- (4-amino-3-methylphenyl) ) -5-fluorobenzothiazole, (5-amino-2,3-fluorophenyl) -6,8-difluoro-7-methyl-4H-1-benzopyran-4-one or their pharmacologically acceptable The determination method according to claim 1, wherein the determination method is a salt. The gene group is a gene group consisting of 25 kinds of genes represented by SEQ ID NOs: 1 to 25. AhR agonist according to claim 1 or 2, which exhibits antitumor activity, a derivative thereof, or a pharmacological agent Of determining the sensitivity of cancer cells to an acceptable salt thereof. The method according to any one of claims 1 to 3, wherein the cancer cells are breast cancer cells, liver cancer cells or ovarian cancer cells. A compound, derivative thereof, or pharmacologically acceptable salt thereof, which is an AhR agonist to which one or more genes selected from the group consisting of 25 genes according to claim 3 are immobilized, and exhibits antitumor activity A microarray for determining the sensitivity of cancer cells to. A kit used for a method for determining sensitivity to an AhR agonist, a compound showing antitumor activity, a derivative thereof, or a pharmacologically acceptable salt thereof, comprising the microarray according to claim 5. A primer set for measuring the expression level of one or more genes selected from the gene group of claim 1. An antibody set for quantifying the amount of intracellular protein of one or more genes selected from the gene group of claim 1.

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