顺铂耐药人宫颈癌HeLa细胞株的建立及其相关耐药微小RNA的表达情况

目的 建立顺铂耐药的人宫颈癌HeLa细胞株,观察其耐药相关微小RNA(miRNA)的表达情况.方法 采用小剂量诱导法建立对顺铂耐药的人宫颈癌细胞株(HeLa/顺铂细胞).比较HeLa细胞株和HeLa/顺铂细胞株的细胞倍增时间、半数抑制浓度(IC50)、耐药指数(RI)、相关耐药基因表达情况及其细胞培养液中肿瘤标志物水平,并分析两种细胞株间异常表达的miRNA表达情况.结果 HeLa/顺铂细胞倍增时间及IC50长于或大于HeLa细胞(均P<0.05),RI值为7.14.HeLa/顺铂细胞培养液癌胚抗原、糖蛋白抗原-125水平均高于HeLa细胞(P<0.05);HeLa顺铂细胞中肺耐药蛋白-1、多药耐药相关蛋白-1及谷胱甘肽S-π的mRNA相对表达量均高于HeLa细胞(P<0.05).HeLa/顺铂细胞miR-29a和miR-34a相对表达量高于HeLa细胞,而miR-99a、miR-92a、miR-155、miR-375、miR-195的相对表达量低于HeLa细胞(均P<0.05).结论 成功建立顺铂耐药HeLa细胞株,其相关耐药miRNA的表达与HeLa细胞株存在差异.

Construction and relevant drug resistance-associated microRNA expression of cisplatin-resistant human cervical cancer cell line HeLa

Objective To construct cisplatin-resistant human cervical cancer cell line HeLa,and to observe the relevant drug resistance-associated microRNA(miRNA) expression of this cell line.Methods Cisplatin-resistant human cervical cancer cell line(HeLa/cisplatin cell) was constructed using low-dose induction.The cell doubling time,half maximal inhibitory concentration(IC50),resistance index(RI),relevant resistance-associated genes expressions and levels of tumor markers in cell culture solution were compared between cell line HeLa and HeLa/cisplatin.The miRNAs expressed abnormally between the two cell lines were analyzed.Results The cell doubling time and IC50 of the HeLa/cisplatin cells were longer or higher than those of the HeLa cells(all P<0.05).The RI of the HeLa/cisplatin cells was 7.14.The carcinoembryonic antigen and carbohydrate antigen-125 levels of cell culture solution were higher in the HeLa/cisplatin cells compared to the HeLa cells(P<0.05).The relative expressions of lung resistance protein 1,multidrug resistance-associated protein 1 and glutathione S-transferaseπ mRNAs were higher in the HeLa/cisplatin cells compared to the HeLa cells(P<0.05).The relative expressions of miR-29a and miR-34a were higher but the relative expressions of miR-99a,miR-92a,miR-155,miR-375 and miR-195 were lower in the HeLa/cisplatin cells compared to the HeLa cell(all P<0.05).Conclusion Cisplatin-resistant cervical cancer cell line HeLa is successfully constructed,and its relevant drug resistance-associated miRNAs expression is different from that of HeLa cell line.