抑制上游转录因子2的表达对胃癌BGC-823细胞增殖和凋亡的影响

Effect of Inhibiting Upstream Transcription Factor 2 Expression on Proliferation and Apoptosis of Gastric Cancer BGC-823 Cells

目的 探讨上游转录因子2(USF2)对人胃癌BGC-823细胞增殖和凋亡的影响。方法 使用Lipofectamine^(TM)3000转染试剂将USF2 siRNA转染至BGC-823细胞(siRNA-USF2组)中,同时设空白对照组和阴性对照组(siRNA-NC组)。实时荧光定量PCR法检测转染后的BGC-823细胞中USF2mRNA的表达。Westernblot实验检测转染后BGC-823细胞中USF2蛋白的表达。CCK-8和平板克隆实验检测每组BGC-823细胞的增殖能力和克隆形成能力。流式细胞术检测各组胃癌细胞的凋亡情况。Western blot实验检测各组BGC-823细胞增殖相关蛋白增殖细胞核抗原(PCNA)及凋亡相关蛋白Bax和Bcl-2的表达水平。结果 与空白对照组和siRNA-NC组相比,siRNA-USF2组细胞USF2 mRNA和蛋白表达显著降低(均P<0.05)。转染后72 h,siRNA-USF2组的吸光度值低于空白对照组(P<0.05)。与空白对照组和siRNA-NC组比较,siRNA-USF2组BGC-823细胞中的克隆数明显较少(P<0.05)。空白对照组、siRNA-NC组和siRNA-USF2组胃癌BGC-823细胞的凋亡率差异具有统计学意义(P<0.05)。与空白对照组和si RNA-NC组比较,si RNA-USF2组BGC-823细胞中PCNA和Bcl-2蛋白表达减少,Bax蛋白表达增加(P<0.05)。结论 抑制USF2表达能够抑制人胃癌细胞的增殖,诱导其凋亡。USF2抑制剂可能在胃癌治疗中具有重要价值。

Objective To investigate the effect of upstream transcription factor 2(USF2) on the proliferation and apoptosis of human gastric cancer BGC-823 cells. Methods Lipofectamine^(TM)3000 transfection reagent was used to transfect USF2 siRNA into BGC-823 cells(siRNA-USF2 group). Blank control and negative control(siRNANC) groups were also prepared. The mRNA and protein expression levels of USF2 in transfected BGC-823 cells were detected by real-time fluorescence quantitative PCR and Western blot, respectively. The proliferation and clone formation abilities of BGC-823 cells in each group were investigated by CCK-8 and plate cloning assay.The apoptosis of gastric cancer cells was examined by flow cytometry. The expression levels of PCNA and apoptosis-related proteins Bax and Bcl-2 in BGC-823 cells were measured by Western blot. Results Compared with those in the blank control and siRNA-NC groups, the mRNA and protein expression levels of USF2 significantly decreased in the siRNA-USF2 group(P<0.05). At 72 h after transfection, the absorbance in the si RNA-USF2 group was lower than that in the blank control group(P<0.05). Compared with that in the blank control and siRNA-NC groups, the number of BGC-823 cell clones significantly decreased in the siRNA-USF2 group(P<0.05). The apoptosis rate of BGC-823 cells significantly differed among the blank control, siRNANC, and siRNA-USF2 groups(P<0.05). Compared with those in the blank control and siRNA-NC groups, the expression of PCNA and Bcl-2 protein decreased and that of Bax protein increased in the siRNA-USF2 group(P<0.05). Conclusion Inhibiting USF2 expression can suppress the proliferation of human gastric cancer cells and induce their apoptosis. USF2 inhibitors may have important value in the treatment of gastric cancer.