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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 inhibits transcription of genes rich in GC sequence promoters. The high expression of GCF2 in hepatoma cell lines such as BEL-7404 may be related to the occurrence and development of liver cancer. In this paper, the expression of GCF2 was silenced by GCF2-siRNA targeting GCF2 to observe the effect of GCF2 on the proliferation and apoptosis of human hepatocellular carcinoma cell line BEL-7404. Methods The experimental cells were divided into two groups: GCF2 siRNA transfected group (transfected with GCF2-siRNA), negative control group (transfected unrelated sequence), lipofectamine 2000 (control group only), normal control group Reagent and siRNA sequences) in 4 groups. GCF2-siRNA was transfected into BEL-7404 cells using Lipofectamine 2000. The expression of GCF2 protein was detected by Western blot 48 h after transfection. The cell counting kit (CCK) -8 and Annexin V / PI-FITC double staining Cytometry was used to detect cell proliferation and apoptosis. Results GCF2-siRNA could effectively down-regulate the expression of GCF2, inhibit the proliferation of BEL-7404 cells and increase the apoptosis rate. Compared with each control group, the differences were statistically significant (P <0.05). Conclusion GCF2 can promote the proliferation and inhibit the apoptosis of hepatocellular carcinoma cell line BEL-7404, suggesting that GCF2 is involved in the occurrence and development of hepatocellular carcinoma through the effects of GCF2 on the proliferation and apoptosis of hepatoma cells.