【摘要】 目的探讨microRNA-21 (miRNA-21)对宫颈癌Hela/DDP耐药的影响及其相关机制。方法实时荧光定量PCR(Real-PCR)检测Hela细胞、Hela/DDP、转染后miR-21-siRNA组及miR-21-neg中miRNA-21的表达量。MTT法检测顺铂(DDP)对Hela/DDP增殖的影响;流式细胞仪分析Hela/DDP细胞周期及凋亡率;Western blot检测顺铂处理过的Hela/DDP中PTEN蛋白的表达。结果在Hela/DDP组中miRNA-21明显高于Hela组,差异具有统计学意义(P<0.05);转染miRNA-21siRNA后,Hela/DDP中miRNA-21明显低于空白对照组及阴性对照组,差异具有统计学意义(P<0.05);与空白对照组及阴性对照组相比,顺铂作用Hela/DDP后,转染miRNA-21 siRNA组细胞增殖抑制率、凋亡率及PTEN蛋白表达水平均升高,并呈现G0/G1期阻滞作用,差异具有统计学意义(P<0.05)。结论 miRNA-21可抑制Hela/DDP细胞增殖、阻滞细胞周期和诱导细胞凋亡,进而提高Hela/DDP细胞对顺铂的敏感性,可能与宫颈癌Hela/DDP细胞中PTEN蛋白合成增加有关。更多还原

【Abstract】 Objective To investigate the effect and mechanisms of miRNA-21 on the resistance of cervical cancer Hela/DDP cells to cisplatin. Methods Real time fluorescence quantitative PCR(Real-PCR) was used to detect the expression of miRNA-21 in Hela cells,Hela/DD,miR-21-siRNA and miR-21-neg after transfection. The effect of cisplatin(DDP) on the proliferation of Hela/DDP was detected by MTT. Apoptosis and cell cycle of Hela/DDP cells were analyzed by flow cytometry. The expression of PTEN protein in Hela/DDP treated with cisplatin was observed by Western blot. Results The relative level of miRNA-21 in Hela/DDP cells was significantly higher than those in Hela cells(P<0.05). The relative level of miRNA-21 in Hela/DDP cells was significantly lower than than those in blank control group and negative control group after transfection with miRNA-21 siRNA(P<0.05). After Hela/DDP cells treatment with cisplatin,the proliferation inhibition rate,the apoptosis rate and the expression level of PTEN protein in the siRNA group were higher than those in the blank control group and the negative control group,and the cells were blocked by the G0/G1 phase(P<0.05). Conclusion The miRNA-21 can inhibit the proliferation of Hela/DDP cells,block cell cycle and induce apoptosis,thereby increasing the sensitivity of Hela/DDP cells to cisplatin,which may be related to the increase of PTEN protein synthesis in cervical cancer Hela/DDP cells.更多还原