人卵巢癌顺铂耐药细胞株的建立及耐药机制研究

目的：建立人卵巢癌顺铂耐药细胞株COC1／DDP，探讨其耐药的机制。方法：采用逐步递增DDP浓度、体外间歇诱导法诱导卵巢癌细胞系COC1，建立卵巢癌顺铂耐药细胞株COC1／DDP；CCK-8试剂盒（Cell Counting Kit-8）检测各种抗癌药对COC1和COC1／DDP细胞增殖的抑制作用，并计算耐药指数；GSH和GSSG试剂盒检测细胞内谷胱甘肽（glutathione，GSH）的水平；高效液相色谱测定细胞内顺铂的含量。结果：历时5个月建立COC1／DDP，对顺铂耐药性稳定，耐药指数为9．44；与COC1比较，COC1／DDP细胞内GSH的水平增高，顺铂含量下降（P〈0．01）。结论：入卵巢癌顺铂耐药细胞株COC1／DDP耐药性稳定，耐药机制可能与GSH／GST-π解毒系统活性增强，减少细胞内顺铂含量有关。

ESTABLISHMENT OF CISPLATIN-RESISTANT HUMAN OVARIAN CANCER CELL LINE COC1/DDP AND INYESTIGATION-OF THE MECHANISM OF RESISTANCE

Objective： To establish a cisplatin-resistant ovarian cancer cell line COC1/DDP, and investigate the mechanism of resistance. Methods： COC1/DDP cell line was established by intermittent-induced method of gradually increasing the concentration of cisplatin to COC1 cell line in vitro.The cytotoxic effect and resistant index of anticancer drugs on COC1/DDP was detected by Cell Counting Kit-8. The level of GSH was determined by GSH and GSSG Kit. The content of cisplatin was detected by HPLC. Results： COC1/DDP cell line was developed after 5 months, with stable resistance to cisplatin, the resistance index was 9.44, Compare to the COC1 cell line , the level of GSH increased but the cisplatin content decreased in COC1/DDP cells （P〈0.01）. Conclusion： The resistance of human cisplatin-resistant ovarian cancer cell line COC1/DDP was stable, its mechanism may be related to decreasing the content of cisplatin by activating the GSH / GST-π detoxification systems.