MicroRNA-1254靶向组蛋白去乙酰化酶7调控胃癌细胞增殖和侵袭

MicroRNA-1254 Regulates Gastric Cancer Cell Proliferation and Invasion Through Targeting Histone Deacetylase 7

[目的]探究microRNA-1254(miR-1254)对胃癌细胞增殖、侵袭的调控作用及分子机制。[方法]使用TargetScan7.1工具预测分析miR-1254对组蛋白去乙酰化酶7(histone deacetylase 7,HDAC7)的靶向作用位点;miR-1254 mimics和miR-1254 inhibitor转染胃癌细胞系SGC-7901,用实时荧光定量PCR检测miR-1254及HDAC7 mRNA表达;CCK-8法检测胃癌细胞增殖活性;Transwell法检测胃癌细胞侵袭能力;蛋白质免疫印迹实验检测STAT3、磷酸化STAT3的表达水平。[结果]MiR-1254可以靶向结合HDAC7 mRNA 3′UTR区域。与对照组相比,转染miR-1254 mimics组miR-1254表达水平升高(t=13.51,P=0.0002),HDAC7 mRNA表达水平显著性降低(t=4.667,P=0.0095),而转染miR-1254 inhibitor组miR-1254表达水平显著性降低(t=10.41,P=0.0005),HDAC7mRNA表达水平显著性升高(t=4.008,P=0.016)。与对照组相比,转染miR-1254 mimics组细胞增殖活性显著性下降(F=30.4,P<0.0001),细胞侵袭能力显著性减弱(t=6.284,P=0.0033);与转染miR-1254 mimics组相比,miR-1254 mimics+HDAC7组细胞增殖活性升高(F=22.16,P=0.0002),且侵袭能力增强(t=5.842,P=0.0043)。转染miR-1254 mimics抑制了STAT3的磷酸化,转染HDAC7表达载体可以减弱miR-1254对STAT3磷酸化的抑制作用。[结论]MiR-1254可以通过靶向HDAC7抑制胃癌细胞STAT3的活化,并抑制细胞的增殖和侵袭。

[Objective]To investigate the effects and molecular mechanism of microRNA-1254(miR-1254)on proliferation and invasion of gastric cancer cells.[Methods]The targeting site of miR-1254 on Histone Deacetylase 7(HDAC7)was analyzed with TargetScan 7.1 online tool.Gastric cancer cells SGC-7901 were transfected with miR-1254 mimics and miR-1254 inhibitors,the expression of miR-1254 and HDAC7 mRNA was analyzed by real-time qPCR.The proliferation rate of gastric cancer cells was measured by CCK-8 assay,cell invasion activities were measured by Transwell assays,expression and phosphorylation levels of STAT3 were detected by immunoblotting assays.The data were statistically analyzed by Graphpad Prim 8.0.[Results]MiR-1254 targeted on the 3′UTR region of HDAC7 mRNA.Compared with control group,the expression of miR-1254 was increased(t=13.51,P=0.0002)and HDAC7 mRNA was decreased(t=4.667,P=0.0095)in miR-1254 mimics group,while the expression of miR-1254 was decreased(t=10.41,P=0.0005)and HDAC7 mRNA was increased(t=4.008,P=0.016)in miR-1254 inhibitor group.Compared with control group,the cell proliferation(F=30.4,P<0.0001)and invasion activities(t=6.284,P=0.0033)were decreased in miR-1254 group;compared with miR-1254 group,the cell proliferation(F=22.16,P=0.0002)and invasion activities(t=5.842,P=0.0043)were decreased in miR-1254+HDAC7 group.Transfection of miR-1254 mimics inhibited STAT3 phosphorylation,and transfection of HDAC7 vectors reduced the inhibition of STAT3 phosphorylation by miR-1254.[Conclusion]MiR-1254 inhibits the activation of STAT3 and the proliferation and invasion of gastric cancer cells through targeting HDAC7.