微小RNA-218对耐顺铂宫颈癌细胞生物学特性影响的研究

Effect of microRNA-218 on the biological characteristics of cisplatin-resistant cervical cancer cells

目的研究和探讨微小RNA-218(miR-218)对耐顺铂宫颈癌细胞的增殖克隆、迁移侵袭能力、药物敏感性等特性的影响。方法以人宫颈癌细胞株SiHa细胞系为亲本株细胞,采用低浓度加量持续诱导法产生耐顺铂SiHa细胞株(耐顺铂SiHa),以慢病毒为载体构建过表达miR-218的耐顺铂细胞株(miR-218/耐顺铂SiHa)。通过荧光定量PCR检测转染后miR-218/耐顺铂SiHa的miR-218表达情况。采用MTT法检测SiHa、耐顺铂SiHa、miR-218/耐顺铂SiHa3组细胞在含顺铂培养液的增殖克隆及顺铂半数有效抑制浓度(IC50)。划痕试验和Transwell法检测3组细胞的迁移和侵袭能力。结果成功构建了耐顺铂SiHa及miR-218/耐顺铂SiHa细胞株,与SiHa及耐顺铂SiHa组比较,miR-218/耐顺铂SiHa组的miR-218表达量显著升高,差异有统计学意义(P<0.05);在含有顺铂培养液中,耐顺铂SiHa组的克隆增殖与IC50显著高于SiHa组与miR-218/耐顺铂SiHa组,差异有统计学意义(P<0.05),SiHa组与miR-218/耐顺铂SiHa组比较,差异无统计学意义(P>0.05),miR-218/耐顺铂SiHa组的划痕愈合率与细胞穿膜次数显著低于SiHa组及耐顺铂SiHa组,且差异有统计学意义(P<0.05)。结论过表达miR-218可以显著抑制宫颈癌细胞增殖与侵袭能力,增强对顺铂的药物敏感性,对耐药病例的治疗提供新的思路。

Objective To investigate the effects of microRNA-218 on the proliferation,migration and drug resistance of cisplatin-resistant cervical cancer cells.Methods Use human cervical cancer cell line SiHa as parent cell line.Cisplatin-resistant SiHa cell line(cisplatin-resistant SiHa)was produced by continuous cisplatin induction at low concentration.Cisplatin-resistant cell line overexpressing miR-218(miR-218/cisplatin-resistant SiHa)was constructed by using lentivirus as vector.The expression of miR-218 in SiHa,cisplatin-resistant SiHa and miR-218/cisplatin-resistant SiHa was detected by real-time PCR.MTT assay was used to detect the proliferation inhibitory concentration(IC 50)of SiHa,cisplatin-resistant SiHa,miR-218/cisplatin-SiHa cells in cisplatin-containing medium.Scratch test and Transwell method were used to detect the migration and invasion ability of the three groups of cells.Results The expression of miR-218 in miR-218/cisplatin-resistant SiHa group was significantly higher than that of SiHa and cisplatin-resistant SiHa group(P<0.05).The expression of miR-218 was significantly higher than that of SiHa group,The cell proliferation and IC 50 of cisplatin-resistant SiHa group were significantly higher than that of SiHa and miR-218/cisplatin-resistant SiHa group(P<0.05).There was no significant difference between SiHa and miR-218/cisplatin-resistant SiHa group(P>0.05),miR-218/cisplatin-resistant SiHa was significantly lower than that of SiHa and cisplatin-resistant SiHa group(P<0.05).Conclusion For cisplatin-resistant cervical cancer cells,overexpression of miR-218 can significantly inhibit its proliferation and invasion,increase the sensitivity to cisplatin,and provide a new idea for the treatment of drug-resistant cases.