两种方法建立的睾丸癌顺铂耐药细胞株的比较.PDF

J South Med Univ, 2015, 35(12): 1755-1759 doi 10.3969/j.issn. 1673-4254.2015.12.18 ·1755 · 技术方法 两种方法建两种方法建立的睾丸癌顺铂耐药细胞株的比较立的睾丸癌顺铂耐药细胞株的比较 李贝贝，董淑英，樊宗兵，吴小祥，巫剑峰，童旭辉 蚌埠医学院药学系药理学教研室，安徽 蚌埠 233030 摘要：目的浓度递增法与大剂量冲击法分别诱导两种小鼠睾丸癌I- 10顺铂耐药细胞株，比较两种细胞株之间形态差异，并检测 耐药相关蛋白MDR 1及P-gp 的表达水平。方法 采用浓度递增法诱导耐药细胞株I- 10/DDPi ，大剂量冲击法的诱导耐药细胞株 I- 10/DDPh ；显微镜下观察细胞形态改变；MTT 法分别检测耐药指数，绘制生长曲线；Western blot 检测耐药相关蛋白MDR 1及 P-gp 的表达；Transwell 法检测细胞侵袭能力的变化。结果 正常睾丸癌I- 10细胞及两种方法建立的相应耐药株均为贴壁生长， 形态饱满，大体呈梭形，上皮样，细胞大小大致相同；I- 10/DDP 细胞大小不同，形态不规则，细胞表面有细长微绒毛类似触角，分 散排列。耐药株I- 10/DDPi 及I- 10/DDPh 对顺铂的耐药指数分别为3.924 和3.099 ，耐药细胞系的倍增时间较正常I- 10 细胞延 长。耐药细胞中耐药相关蛋白MDR 1及P-gp 表达显著高于正常I- 10 细胞，其中I- 10/DDPi 增加更为明显。耐药细胞株侵袭力 增强，其中I- 10/DDPh 增加更为明显。结论 采用两种方法均可成功建立的耐顺铂睾丸癌细胞株，且大剂量冲击法建立的耐顺 铂细胞株I- 10/DDPh 更接近于临床中的肿瘤耐药细胞。 关键词：睾丸癌；顺铂；耐药 Comparison of cisplatin-resistant testicular cancer cell lines established by two methods LI Beibei, DONG Shuying, FAN Zongbing, WU Xiaoxiang, WU Jianfeng, TONG Xuhui Department of Pharmacology, Faculty of Pharmacy, Bengbu Medical College, Bengbu 233030, China Abstract: Objective To compare the biological behaviors of two drug-resistant testicular cancer cell lines established by different methods. Methods Drug-resistance was induced in testicular cancer cell lines exposure of the cells to increasing concentrations of or a high dose of cisplatin (I-10/DDPi and I-10/DDPh cell lines, respectively). The morphological characteristics of the two cell lines were observed microscopically. The resistance index of the cells was determined with MTT assay, and the cell growth curves were drawn. The cellular expression of resistance-associated proteins MDR1 and P-gp was detected by Western blotting. The cell invasion ability was assessed with Transwell assay. Results Normal testicular cancer cell line I-10 and the two resistant cell lines all showed an adherent