RNA干扰GCF2基因沉默对人肝癌BEL-7404细胞侵袭能力的影响

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

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。

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 as- sessed 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.