南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (12): 1755-.

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两种方法建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   

  • 出版日期:2015-12-20 发布日期:2015-12-20

Comparison of cisplatin-resistant testicular cancer cell lines established by two methods

  • Online:2015-12-20 Published:2015-12-20

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MDR1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MDR1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MDR1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。

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 growth pattern. Compared with I-10 cells, I-10/DDP cells
exhibited slightly heterogenous cell sizes, irregular shapes, the presence of microvilli tentacles on the cell surface, and a
scattered arrangement. The cisplatin resistance index of I-10/DDPi and I-10/DDPh cells were 3.924 and 3.099, respectively.
Compared with I-10, the drug-resistant cell lines showed extended doubling time with increased expressions of MDR1 and
P-gp and increased cell invasiveness, which was especially obvious in I-10/DDPi cells. Conclusion Both increasing dose
exposure and high-dose exposure to cisplatin can induce cisplatin resistance in testicular cancer cells, and the resistant cells
established by the latter method better mimics clinical drug-resistant tumor cells.