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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。
Objective To establish a cisplatin-resistant human cervical cancer HeLa cell line and observe its expression of drug-resistant microRNA (miRNA). Methods Cisplatin-resistant human cervical cancer cell lines (HeLa / cisplatin cells) were established by low-dose induction. The cell doubling time, IC50, RI, the expression of related drug resistance genes and the levels of tumor markers in HeLa and HeLa / cisplatin cell lines were compared and analyzed Expression of miRNAs abnormally expressed in cell lines. Results HeLa / Cisplatin cell doubling time and IC50 were longer or longer than HeLa cells (all P <0.05), RI value was 7.14. HeLa / cisplatin cell culture fluid carcinoembryonic antigen, glycoprotein antigen -125 levels were higher than HeLa cells (P <0.05); HeLa cisplatin cells lung resistance protein-1, multidrug resistance protein-1 and Valley The relative expression of S-π mRNA in HepG2 cells was higher than that in HeLa cells (P <0.05). The relative expression levels of miR-29a and miR-34a in HeLa / cisplatin cells were higher than those in HeLa cells, while the relative expression levels of miR-99a, miR-92a, miR-155, miR- P <0.05). Conclusion The successful construction of cisplatin-resistant HeLa cell lines, the expression of related drug-resistant miRNA HeLa cell lines there are differences.