白藜芦醇通过下调microRNA-151表达抑制肝癌细胞株HepG2细胞活性的研究

Inhibition of hepatocellular carcinoma HepG2 cell line through down-regulation of expression of microRNA-151 by resveratrol

目的研究白藜芦醇(Res)对肝癌细胞株HepG2宿主基因黏着斑激酶(FAK)和内含子miR-151表达的影响,探讨其抗肿瘤的可能作用机制。方法分别以50、100、150、200μmol/LRes作用于体外培养的HepG2细胞24、48和72h。CCK-8法检测细胞增殖;Real-Time PCR检测细胞FAK mRNA和miR-151表达。利用miR-151模拟物转染获得miR-15过表达HepG细胞株(miR-151过表达组),流式细胞仪和Hoechst33342染色观察过表达miR-151对HepG2细胞周期及凋亡的影响。以转染模拟物突变体细胞作为阴性对照,未转染细胞作为空白对照。结果 Res对体外培养HepG2细胞的增殖活性及FAK mRNA和miR-151表达具有显著抑制作用,并在一定范围内呈现浓度和时间依赖性。miR-151过表达组HepG细胞miR-151表达明显高于阴性对照组和空白对照组(P<0.001);流式细胞仪检测和Hoechst33342染色观察结果显示,miR-15过表达使HepG细胞G0/G1期缩短,细胞周期进程加快,并能抑制细胞凋亡。结论 Res可能通过下调miR-151表达,抑制肝癌细胞HepG2增殖并诱导凋亡。

Objective To investigate the effects of resveratrol （Res） on expression of host gene focal adhesion kinase （FAK） and its intronic microRNA （miR-151） in hepatocellular carcinoma HepG2 cells, and explore the possible mechanism of its anti-tumor effect. Methods HepG2 cells cultured in vitro were treated with 50, 100, 150 and 200 μmol/L Res for 24, 48 and 72 h, respectively. CCK-8 assay was employed to examine the cell proliferation, and the expression of FAK mRNA and miR-151 was detected by Real-Time PCR. Transfection with miR-151 mimics was conducted, and HepG cells with miR-15 overexpression were obtained （miR-151 overexpression group）. Flow cytometry and Hoechst 33342 staining were adopted to detect the effects of miR-15 overexpression on cell cycle and apoptosis of HepG2 cells. Cells transfected with mimics mutants and those without transfection were served as negative controls and blank controls, respectively. Results Res significantly inhibited the cell proliferation and decrease the expression of FAK mRNA and miR-151 of HepG2 cells in a concentration and time-dependent manner to some degree. The expression of miR-151 of HepG2 cells in miR-151 overexpression group was significantly higher than that in negative control group and blank control group （P〈0.001）. Detection by flow cytometry and Hoechst 33342 staining revealed that miR-151 overexpression shortened G0/G1 phase, fastened cell cycle progression and inhibited cell apoptosis of HepG2 cells. Conclusion Res may inhibit proliferation and induce apoptosis of hepatocellular carcinoma HepG2 cells through down-regulation of expression of miR-151.