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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 investigate the effect of silencing GCF2 gene expression on invasiveness of human hepatocellular carcinoma BEL-7404 cells. Methods The experiment was divided into GCF2-siRNA group, NC-siRNA group and MOCK group. GCF2-siRNA group was transfected with GCF2-siRNA targeting GCF2 gene and negative control group was transfected with NC-siRNA and MOCK group Transfect any sequence. The GCF2-siRNA targeting GCF2 gene was transiently transfected into BEL-7404 cells to silence the expression of GCF2. The cell proliferation activity was detected by CCK-8 assay. The invasion ability was detected by in vitro invasion assay. The expression of matrix metalloproteinase (MMP) -9 and GCF2 target gene MMP-23 protein expression. Results GCF2-siRNA transfected GCF2 down-regulated the expression of GCF2, which led to the obstruction of proliferation of BEL-7404. The number of transmembrane cells in GCF2-siRNA group was significantly decreased compared with that in NC-siRNA group and non-transfected group Membrane cells were 41.5 ± 0.95, 61.3 ± 1.57, 64.5 ± 1.65, P <0.05). The protein expressions of MMP-9 and MMP-23 were significantly decreased compared with the two control groups (P <0.05). Conclusion Down-regulation of GCF2 expression can inhibit the invasion ability of BEL-7404, suggesting that GCF2 may promote the invasion of hepatocellular carcinoma cells and may be an important pathway involved in the development of hepatocellular carcinoma.