MiR-218在宫颈癌组织中的表达及对HeLa细胞凋亡及转移的影响

Expression of Micro-RNA 218 in Cervical Cancer and Its Effect on Proliferation,Apoptosis and Invasion of HeLa Cells

目的探讨miR-218在宫颈癌组织中的表达,以及对宫颈癌HeLa细胞凋亡及迁移的影响。方法采用qRT-PCR检测23例正常子宫颈组织和114例宫颈癌组织中miR-218mRNA的表达,分析与临床病理特征的关系;HeLa细胞分为3组:对照组(细胞不转染)、转染空脂质体阴性对照组(NC组)和转染miR-218类似物(miR-218M组)。MTT检测细胞增殖;流式检测细胞凋亡;划痕实验和Transwell实验检测细胞迁移;qRT-PCR和Western blot分别检测Bcl-2、Bax、NF-κB和E-cadherin mRNA和蛋白表达。结果 MiR-218mRNA在宫颈癌组织中表达下调,在宫颈癌不同病理类型、分期分级、有无淋巴结转移及间质浸润深度间表达差异有统计学意义(P<0.01);采用miR-218类似物转染宫颈癌HeLa细胞,细胞增殖减少,凋亡增加,侵袭转移能力减弱。qRT-PCR和Western blot检测结果显示,转染miR-218类似物,HeLa细胞Bcl-2mRNA和蛋白表达水平下调,而Bax mRNA和蛋白表达水平上调,E-cadherin mRNA和蛋白表达上调,NF-κB mRNA和蛋白表达下调。结论 MiR-218低表达可能与宫颈癌的病理分级分期等生物学行为有关,上调miR-218表达可抑制细胞增殖,促进细胞凋亡,抑制细胞侵袭转移。

Objective To investigate the expression and clinical significance of microRNA-218 （miR-218） in human cervical cancer and the effects of miR-218 on proliferation, cell apoptosis and invasion of HeLa cells. Methods QRT-PCR was used to detect the expression of miR-218 in 23 cases of normal cervical tissues and 114 cases of cervical cancer, and the relationship between the expression and the clinicopathological features was analyzed HeLa cells were devided into three groups., non transfection （control group）, transfected with empty liposomes negative control goup （NC group）, transfected with miR-218 mimic （miR-218M group）. The cell growth inhibiting ratio of HeLa cells was assessed by MTT assay. Fluorescence activated cell sorting was used to measure cell apoptosis. Changes of cell migration ability were detected by wound healing test and Transwell assay. QRT- PCR and Western blot were used to detect the expression of Bcl-2, Bax, NF-κB and E-cadherin, respectively. Results The expression of miR-218 in cervical cancer was clown regulated, and there were significant differences in the different pathological types, stages, lymph node metastasis and interstitial infiltration in cervical cancer tissues （P〈0.01）; After being transfected with miR-218 mimic, the proliferation of HeLa cells was significantly inhibited. The ability of invasion was decreased. QRT-PCR and Western blot showed that after being transfected with miR- 218 mimic, the expression levels of Bcl-2 mRNA and protein were down-regulated and Bax mRNA and protein expression levels were increased, E-cadherin mRNA and protein expression were up-regulated, but NF-IcB mRNA and protein expression were down-regulated. Conclusion The low-expression of miR-218 is correlated with the poor clinicopathological features in human cervical cancer. MiR-218 overexpression reduces cancer cell proliferation and induces apoptosis and inhibits cell migration, suggesting that miR-218 may play a key role in the progression of human cervical cancer.