体外培养人卵巢癌细胞株SKOV-3、SKOV-3/DDP并探讨卵巢癌顺铂耐药机制

Investigate the molecular mechanism of the cisplatin-resistant with human epithelial ovarian cancer cell line skov-3 and skov-3/DDP cultured in vitro

目的:通过对人卵巢癌细胞株SKOV-3、SKOV-3/DDP体外培养,探讨卵巢癌顺铂耐药机制。方法:用四甲基偶氮唑蓝(MTT)法、流式细胞仪测定SKOV-3、SKOV-3/DDP细胞顺铂耐药指数及细胞周期变化,并分析细胞内Bcl-2蛋白及细胞膜上Fas、Fas-L表达情况。结果:相对于SKOV-3细胞;SKOV-3/DDP细胞顺铂耐药指数约为4倍,其细胞周期主要分布于G0/G1期、S期,细胞内Bcl-2蛋白表达及细胞膜Fas、Fas-L水平明显升高。结论:细胞主要分布于潜伏期、细胞内凋亡抑制蛋白Bcl-2水平升高及细胞膜Fas、Fas-L表达增加,可能是卵巢癌顺铂耐药细胞株SKOV-3/DDP顺铂耐药的部分机理。

Objective:To investigate the molecular mechanism of the cisplatin-resistant with human epithelial ovarian cancer cell line skov-3 and skov-3/DDP cultured.Methods:The inhibition of skov-3 and skov-3/DDP cells growth were measured by methyl thiazolyl tetrazolium(MTT) rapid photocolorimetric assay,respectively. The resistance index was calculated by skov-3/DDP(A)/ skov-3(A). Cell cycle of two cell lines was accounted by flow cytometry. In addition, expression of Fas、Fas-L in cell membrane and intracellular Bcl-2 protein were analyzed by flow cytometry.Results:In contrast to skov-3 cell , the resistance index of skov-3/DDP cell was nearly four fold.Cell cycle of skov-3/DDP cell line mainly distributed G0/G1 and S phase , but those of skov-3 cell line principally distributed S phase.expression of Fas、Fas-L in cell membrane and intracellular Bcl-2 protein of skov-3/DDP cell line obviously enhanced more than those of skov-3 cell line.Conclusion:Dormient period cell increased and expression of Fas、Fas-L and Bcl-2 protein enhanced may be the partial mechanisms of the cisplatin-resistant of skov-3/DDP cell line.