RNA干扰GCF2基因沉默对人肝癌BEL-7404细胞侵袭能力的影响

目的 探讨沉默GCF2基因表达对人肝癌BEL-7404细胞侵袭能力的影响.方法 实验分为GCF2-siRNA组、NC-siRNA组及MOCK组,GCF2-siRNA组转染靶向GCF2基因的RNA干扰序列GCF2-siRNA,阴性对照组转染阴性对照序列NC-siRNA,MOCK组不转染任何序列.将靶向GCF2基因的RNA干扰序列 (GCF2-siRNA) 瞬时转染BEL-7404细胞以沉默GCF2表达,采用CCK-8法检测细胞增殖活性,体外侵袭实验检测细胞侵袭能力,Western blot检测基质金属蛋白酶(MMP)-9及GCF2靶基因MMP-23蛋白表达.结果 转染GCF2-siRNA使GCF2表达下调,后者导致BEL-7404增殖受阻;GCF2-siRNA组的穿膜细胞数较阴性对照组(NC-siRNA)及未转染组(MOCK)明显减少(穿膜细胞数分别为41.5±0.95、61.3±1.57、64.5±1.65,P<0.05);MMP-9和MMP-23蛋白表达较两个对照组明显下降(P<0.05).结论 下调GCF2表达能抑制BEL-7404侵袭能力, 提示GCF2促进肝癌细胞的侵袭可能是其参与肝癌发展进程的重要途径.

Effect of RNAi silencing GCF2 on the invasion of human hepatocellular carcinoma cell line BEL-7404

Objective To observe the effect of GCF2 on invasion of human hepatocellular carcinoma cell line BEL-7404.Methods The siRNA targeting GCF2 mRNA (named as GCF2-siRNA) was transiently transfected into BEL-7404 cells.Forty-eight hours after transfection, cell proliferation was determined by cell counting Kit-8, and cell invasion ability was determined by Tran-swell invasion assay.MMP-9 and MMP-23 protein expression was assessed by western blotting.Results GCF2-siRNA effectively down-regulated GCF2 expression.Compared with the negative control group and MOCK group, cell proliferation and invasion abilities in siRNA-GCF2 group were remarkably suppressed.Meanwhile, MMP-9 and MMP-23 protein expression was also obviously reduced.Conclusion Down-regulation of GCF2 by siRNA suppresses cell invasion abilities of BEL-7404, which suggests that GCF2 plays a crucial role in the progression of HCC by affecting cell invasion.