上调microRNA-150表达对肝癌SMMC7721细胞增殖和凋亡的影响

目的 探讨上调microRNA- 150 (miR- 150)表达对肝癌SMMC7721细胞增殖和凋亡的影响及其可能机制.方法 SMMC7721细胞分成miR-150感染组(加入pGIPZ-miR-150慢病毒表达载体)、阴性对照组(加入只含GFP的慢病毒载体)和空白对照组(不转染).采用荧光定量PCR检测miR-150的表达情况,Western blotting检测靶基因c-Myb蛋白的表达,CCK-8法检测细胞增殖能力,流式细胞仪检测细胞凋亡情况.结果 荧光定量PCR结果显示,转染miR-150pGIPZ后,SMMC7721细胞miR-150表达量(2.48±0.15)较阴性对照组(0.81±0.09)增加了2倍(p＜0.01).CCK-8法检测结果显示,与阴性对照组和空白对照组比较,miR-150组的细胞增殖率显著降低(P＜0.05).Western blotting检测结果表明,miR-150组细胞c-Myb蛋白表达(0.31±0.07)与空白对照组(0.80±0.09)及阴性对照组(0.81±0.08)相比明显减少(P＜0.0l).流式细胞仪检测结果显示,miR-150组细胞凋亡率(19.36％±1.78％)与阴性对照组(5.12％±0.54％)及空白对照组(4.68％±0.35％)相比明显增加(P＜0.01).结论 上调miR-150表达可通过降低靶基因c-Myb的表达,抑制细胞增殖,诱导细胞凋亡,miR-150可能成为肝癌靶向治疗新的靶基因.

Effect of microRNA-150 up-regulation on proliferation and apoptosis of hepatocellular carcinoma cell line SMMC-7721

Objective To investigate the effects of microRNA150(miR-150) up-regulation on proliferation and apoptosis of hepatocellular carcinoma cell line SMMC7721,and the potential mechanism thereof.Methods The SMMC7721 cells were divided into miR-150 modulated group,negative control group,and blank control group.Chemically synthesized hsa-miR-150,according to the maturity sequence of hsa-miR-150,was inserted into pGIPZ vector.SMMC7721 cells in miR-150 modulated group and negative control were transfected with miR-150pGIPZ and miR-control respectively,while those in blank control group without transfection.The expression of miR-150 was determined by real-time fluorescence quantitative PCR.The cell proliferation ability was determined by Cell Counting Kit-8(CCK-8) assay,and cell apoptosis was assayed by flow cytometry.The protein expression of c-Myb was measured by Western blotting.Results The miR-150 expression increased by 7 folds in cells after being transfected with miR-150 than those in negative control group and blank control group.CCK-8 assay showed that the cellular growth rate of SMMC7721 cells in miR-150pGIPZ group was significantly lower than those in negative control group and blank control group(P<0.05).The apoptotic rate in miR-150pGIPZ group,negative control group and blank control group was(19.36±1.78)%,(5.12±0.54)% and(4.68±0.35)%,respectively.The expression level of c-Myb protein in negative control group,blank control group and miR-150pGIPZ group was 0.31±0.07,0.80±0.09 and 0.81±0.08,respectively.Conclusion The results indicate that up-regulation of miR-150 may suppress the cellular proliferation and induce cell apoptosis in SMMC7721 cells,at least in part,through regulating the expression level of target gene c-Myb,thus implying that miR-150 might serve as a novel molecular target for treatment of HCC.