microRNA-99b表达调控对宫颈癌HeLa细胞增殖和凋亡的作用

目的:探讨上调microRNA-99b(miR-99b)表达对宫颈癌HeLa细胞增殖和凋亡的作用及可能的机制。方法:利用转染试剂将Pre-miR-hsa-miR-99b miRNA Precursor转染宫颈癌HeLa细胞,荧光实时定量PCR方法检测miR-99b表达,四甲基偶氮唑蓝(MTT)法检测细胞增殖能力,流式细胞仪检测细胞凋亡率,Western blot检测其靶基因CDC-25A蛋白的表达。结果:转染Pre-miR-hsa-miR-99b miRNA Precursor的HeLa细胞,miR-99b表达增高86.45倍,细胞增殖能力下降,细胞凋亡率增加,同时细胞CDC-25A蛋白表达降低。结论:上调miR-99b表达可通过降低靶基因CDC-25A表达,抑制细胞增殖,诱导细胞凋亡,miR-99b可能成为治疗宫颈癌的靶基因。

Effects of microRNA-99b modulation on proliferation and apoptosis of cervical cancer HeLa cells

Objective:To investigate the effects of microRNA-99b(miR-99b)up-regulated on proliferation and apoptosis of HeLa cells and its possible mechanism.Methods:Pre-miR-hsa-miR-99b miRNA Precursor was transfected into HeLa cells with transfection agent.The miR-99b expression was detected by real time PCR.The cellular growth activity was assayed by MTT assay,and the apoptosis was tested by flow cytometry.The protein expression of CDC-25A were measured by Western blot.Results:The miR-99b expression in Pre-miR-hsa-miR-99b miRNA Precursor transfected cells increased 86.45 fold.The cellular growth activity decreased,and the cell apoptosis increased.Furthermore,the protein expression of CDC-25A decreased in Pre-miR-hsa-miR-99b miRNA Precursor transfected cells.Conclusion:The miR-99b up-regulation can suppress the cellular proliferation and induce apoptosis in HeLa cells by decreased target genes CDC-25A expression.The miR-99b may be a target gene for cervical carcinoma treatment.