siRNA沉默GC结合因子2表达对BEL-7404细胞增殖和凋亡的影响

Effects of GCF2 silenced by siRNA on the proliferation and apoptosis of human hepatocellular carcinoma cell line BEL-7404

目的 GC结合因子2(GC binding factor 2,GCF2)是一种转录抑制因子,能抑制富含GC序列启动子的基因转录。GCF2在BEL-7404等肝癌细胞株的高表达可能与肝癌的发生及发展有关。文中通过靶向GCF2的GCF2-siRNA序列沉默GCF2表达,观察GCF2对人肝癌细胞株BEL-7404增殖及凋亡的影响。方法将实验细胞分为GCF2 siRNA转染组(转染GCF2-siRNA)、阴性对照组(转染无关序列)、脂质体对照组(只加Lipofectamine 2000)、常规培养对照组(不加转染试剂和siRNA序列)共4组。使用Lipofectamine 2000将GCF2-siRNA转染BEL-7404细胞,Western blot检测转染后48 h GCF2蛋白表达水平,用细胞计数试剂盒(cell counting kit,CCK)-8和AnnexinV/PI-FITC双染流式细胞术检测细胞增殖和凋亡变化。结果 GCF2-siRNA可有效下调GCF2表达、抑制BEL-7404细胞的增殖,细胞凋亡率增加。与各对照组相比,差异均有统计学意义(P<0.05)。结论 GCF2可促进肝癌细胞株BEL-7404的增殖,抑制该细胞凋亡,提示GCF2通过对肝癌细胞增殖和凋亡的影响参与肝癌的发生、发展。

Objective GC-binding factor 2 （ GCF2 ） is a transcriptional repressor that represses the transcription of the genes rich in GC sequences in their promoters. The overexpression of the GCF2 protein in human hepatoeellular carcinoma （ HCC ） cell lines, such as BEL-7404, is involved in the occurrence and progression of HCC. The aim of this study is to observe the effect of transient transfection with siRNA targeting GCF2 on the proliferation and apoptosis of BEL-7404 cells. Methods The siRNA tar- geting GCF2 was transiently transfected into BEL-7404 cells by lipofectamine 2000. The expression of the GCF2 protein was detected by Western blot at 48 hours after transfection. The proliferation and apoptosis of BEL-7404 cells were determined by cell counting kit-8 and Annexin V-FITC flow cytometry, respectively. Results Transfection of GCF2 siRNA effectively down-regulated the ex-pression of GCF2, inhibited the proliferation and promoted the apoptosis of BEL-7404 cells （P 〈 0.05）. Conclusion GCF2 can promote the proliferation and inhibit the apoptosis of BEL-7404 cells, which suggests that GCF2 plays a vital role in the occurrence and progression of HCC.