人宫颈癌顺铂耐药细胞系SiHa/cDDP的建立

目的建立人宫颈癌SiHa细胞对顺铂(cDDP)的耐药细胞系并分析其生物学特性。方法用化疗药物cDDP与人宫颈癌细胞系SiHa共培养,逐步增加培养液中cDDP浓度以诱导细胞产生耐药性,建立SiHa细胞对cDDP的耐药细胞系。利用MTT法绘制细胞生长曲线,计算细胞倍增时间并检测肿瘤细胞耐药指数(RI)。免疫细胞化学法检测细胞P-糖蛋白(P-gp)、胎盘型谷胱甘肽-S-转移酶(GST-π)、拓扑异构酶Ⅱ(TopoⅡ)蛋白的表达。结果成功建立耐药细胞系SiHa/cDDP(耐cDDP 2μg/mL),倍增时间(45.82±3.69)h,对cDDP的RI为16.131,对多种化疗药物有不同程度交叉耐药。耐药细胞P-gp、GST-π表达较亲代细胞增多(P<0.01),TopoⅡ表达与亲代细胞间差异无统计学意义。结论本研究建立了人宫颈癌顺铂耐药细胞系SiHa/cDDP,为肿瘤细胞体外研究提供了理想的实验模型。

Establishment of a cisplatin-resistant human cervical cancer cell line

Objective To establish a cisplatin(cDDP)-resistant human cervical cancer cell line named SiHa/cDDP and researched its biological characteristics.Methods The development of cDDP resistance in SiHa cell line was induced by continuously stepwise exposure of the cells to cDDP.Cell growth curve,doubling time and resistance index(RI) were evaluated by MTT analysis.The expression of P-gp,GST-π,TopoⅡ were assessed with immunocytochemical method.Results SiHa/cDDP cell line was successfully established which had stable growth,subculture,frozen reservation and resuscitation in the concentration of 2 μg/ml cDDP,doubling time was(45.82±3.69) h and RI to cDDP was 16.131.It also showed different degrees of cross-resistance to several anticancer drugs.As compared to parental SiHa,SiHa/cDDP had over-expression in P-gp and GST-π(P<0.01),but TopoⅡ showed no statistically significant differences.Conclusion We successfully established the cDDP-resistant human cervical cancer cell line SiHa/cDDP,which may provide ideal experimental model for research of human cervical carcinoma.