Sakamoto M, Kondo A, Kawasaki K, Goto T, Sakamoto H, Miyake K, Koyamatsu Y, Akiya T, Iwabuchi H, Muroya T, Ochiai K, Tanaka T, Kikuchi Y, Tenjin Y
Dept. of Gynecology, Sasaki Institute Kyoundo Hospital.
Hum Cell. 2001 Dec;14(4):305-15.
Gene expression profiles were analyzed by using cDNA microarray for a cisplatin-sensitive cell line (KF), and three- and thirty-fold cisplatin-resistant ovarian cancer cell lines (KFr and KFrP200) both showing no p53 mutation within exon 5, 6, 7, 8 and no pglycoprotein overexpression. Expression of GST-pi mRNA increased as the level of resistance to cisplatin became high. Microarray analysis revealed that DNA repair associated genes, i.e., XRCC5, XRCC6, ERCC5, hMLH1 were over-expressed in three-fold cisplatin-resistant cell line, KFr as compared to cisplatin-sensitive parental cell line, KF. Apoptosis inhibitors, i.e., IGFR type I and II were over-expressed, and apoptosis inducer, i.e., caspase 3 and BAK were underexpressed in highly cisplatin-resistant cell line, KFrP200 as compared to KFr. As for clinical cases, cDNA microarray was used to compare gene expression profiles directly between two groups, i.e., the chemotherapy (CAP) sensitive group (n = 2) and the resistant group (n = 2). Six genes such as beta tubulin, high-mobility group (nonhistone chromosomal) protein 1, connective tissue growth factor, insulin-like growth factor binding protein 2, alpha tubulin, and RAS-related gene were overexpressed in CAP therapy resistance group, whereas seven genes such as CD9 antigen, alpha-2-macroglobulin, caveolin 2, interleukin 1 receptor antagonist, Rho GTPase activating protein 1, reticulon 3, cyclin-dependent kinase 10, keratin 7 were underexpressed in CAP therapy resistance group. By increasing clinical case number and gene number of microarray to be used in the analysis of expression profile of gene cluster affecting anticancer drug resistance and sensitivity of the ovarian cancer, it would be possible to apply microarray analysis to personalization of chemotherapy such as selection of effective chemotherapy protocol and prediction of therapeutic effect in the near future.
利用cDNA微阵列分析了顺铂敏感细胞系（KF）、三倍和三十倍顺铂耐药卵巢癌细胞系（KFr和KFrP200）的基因表达谱，这两种耐药细胞系在第5、6、7、8外显子均无p53突变，且无P-糖蛋白过表达。随着对顺铂耐药水平的升高，谷胱甘肽S-转移酶π（GST-pi）mRNA的表达增加。微阵列分析显示，与顺铂敏感的亲本细胞系KF相比，三倍顺铂耐药细胞系KFr中与DNA修复相关的基因，即X射线修复交叉互补基因5（XRCC5）、X射线修复交叉互补基因6（XRCC6）、切除修复交叉互补基因5（ERCC5）、人错配修复蛋白1（hMLH1）过表达。与KFr相比，在高度顺铂耐药细胞系KFrP200中，凋亡抑制剂，即Ⅰ型和Ⅱ型胰岛素样生长因子受体（IGFR）过表达，而凋亡诱导剂，即半胱天冬酶3（caspase 3）和BAK蛋白低表达。对于临床病例，使用cDNA微阵列直接比较两组之间的基因表达谱，即化疗（CAP）敏感组（n = 2）和耐药组（n = 2）。在CAP治疗耐药组中，β微管蛋白、高迁移率族（非组蛋白染色体）蛋白1、结缔组织生长因子、胰岛素样生长因子结合蛋白2、α微管蛋白和RAS相关基因等6个基因过表达，而CD9抗原、α-2-巨球蛋白、小窝蛋白2、白细胞介素1受体拮抗剂、Rho GTP酶激活蛋白1、网质蛋白3、细胞周期蛋白依赖性激酶10、角蛋白7等7个基因在CAP治疗耐药组中低表达。通过增加用于分析影响卵巢癌抗癌药物耐药性和敏感性的基因簇表达谱的临床病例数量和微阵列基因数量，在不久的将来将微阵列分析应用于化疗个体化，如选择有效的化疗方案和预测治疗效果将成为可能。
