胃癌顺铂耐药细胞株的建立和基因表达谱分析  

Establishment of DDP-resistant gastric cancer cell line SGC7901 and gene profile analysis

作　　者：李大卫[1] 彭志海[1] 吴晴[2] 王静珏 杨海燕[2] 

机构地区：[1]上海交通大学附属第一人民医院普外科,200080 [2]上海交通大学附属第一人民医院肿瘤科,200080

出　　处：《中华实验外科杂志》2009年第1期31-33,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：上海市自然科学基金资助项目（03ZR14077）

摘　　要：目的建立人胃癌顺铂耐药细胞株，探讨其基因表达谱与顺铂耐药的相关性。方法采用浓度梯度递增法诱导胃癌SGC7901细胞株，建立对顺铂耐药的SGC7901／DDP细胞株。应用Affymetrix基因芯片HG-U133 Plus2．0筛选SGC7901和SGC7901／DDP的耐药相关的差异基因。结果建立可耐受1．0mr,／LJ顷铂浓度的耐药株SGC7901／DDP，耐药指数22．85。应用电镜观察其形态学变化，细胞计数法绘制生长曲线，噻唑蓝（MYr）检测检测药物敏感性，流式细胞术分析细胞周期。利用基因芯片筛选得到在耐药细胞株和敏感株中表达差异大于10倍的基因共107条，其中JNK1和JNK2为与耐药相关的明显上调基因。Western blot证实JNK的磷酸化活性形式P-JNK在耐药株中表达明显升高。SP600125抑制P-JNK表达后耐药蛋白P-gP表达明显减低。结论高通量的基因芯片筛选得到大量有意义的与顺铂耐药相关的差异基因，P-JNK可成为逆转耐药的新的靶点。Objective To establish cisplatin （DDP）-resistant human gastric cancer cell line and investigate the relationship between differential gene profile and DDP resistance. Methods A DDP-resistant human gastric cancer cell line SGC7901/DDP was established by increasing dose of DDP up to 1.0 mg/L and the resistance index （RI） was 22.85. The multidrug resistance （MDR） phenotype was tested by detecting cell morphology using electron microscopy, drug sensitivity using MTT array, and cell cycle using flow cytometry. Results 107 genes were identified,which were differentially expressed by more than 10 times between SGC7901 and SGC7901/DDP cell lines using Affymetrix gene chip HG-U133 Plus 2.0, among which JNK1 and JNK2 were significantly up-regulated in SGC7901/DDP cell line and the active phosphorylation type of JNK （P-JNK） was verified by Western blot. The expression of MDR protein P-gp was inhibited after the blocking of P-JNK by SP600125. Conclusion Numerous genes relating with DDP- resistance were screened out by through-out gene chip and P-JNK could be the novel target gene for reversing MDR.

关 键 词：胃肿瘤 基因芯片 多药耐药 

分 类 号：R735.2[医药卫生—肿瘤]