沉默SEMG1蛋白对精原细胞GC-1细胞周期及凋亡的影响

Effects of silencing the SEMG1 protein on the cycle and apoptosis of GC-1 spg cells

目的:探究沉默生精蛋白Ⅰ(SEMG1)对精原细胞系GC-1细胞周期及凋亡的影响。方法:利用脂质体转染法将SEMG1小干扰RNA(siRNA)转染精原细胞株GC-1,以空白组(空白对照组)和非特异性siRNA转染组(阴性对照组)作为对照;使用Western印迹法检测各组细胞SEMG1蛋白的表达水平,流式细胞术分析各组细胞的凋亡率及细胞周期。结果:与空白对照组和非特异性siRNA转染组相比,siRNA-SEMG1(si-SEMG1)转染组细胞的SEMG1蛋白表达水平(2.51±0.13,2.50±0.12 vs 1.80±0.05)明显降低(P<0.01);空白对照组、阴性对照组和siRNA-SEMG1转染组细胞的细胞周期无明显差异(P>0.05),但siRNA-SEMG1转染组细胞凋亡率达到(6.77±0.15)%,明显高于空白对照组(0.70±0.06)%和阴性对照组(0.8±0.06)%(P<0.01)。此外,Western印迹结果显示,siRNA-SEMG1转染组细胞的caspase-3蛋白表达水平显著上升(P<0.05),而Bcl-2蛋白表达水平明显下降(P<0.05)。结论:siRNA-SEMG1能有效沉默精原细胞GC-1中SEMG1蛋白的表达,并最终促进GC1-细胞凋亡。

Objective:To investigate the effects of silencing the semenogelin 1(SEMG1)protein on the cycle and apoptosis of the spermatogonia germ cell line(GC-1 spg).Methods:SEMG1-specific siRNA was transfected into GC-1 spg cells by lipofectamine 2000(the siRNA-SEMG1 group),the relative expression levels of the SEMG1 protein in the GC-1 spg cells of the siRNA-SEMG1,blank control and negative control groups were detected by Western blot,and the apoptosis and cycle of the cells in different groups were determined by flow cytometry.Results:The expression of the SEMG1 protein in the GC-1 spg cells was dramatically decreased in the siRNA-SEMG1 group compared with those in the blank and negative control groups(1.80±0.05 vs 2.51±0.13 and 2.50±0.12,P<0.01),but the apoptosis rate was remarkably higher in the former than in the latter two groups([6.77±0.15]%vs[0.70±0.06]%and[0.8±0.06]%,P<0.01).No statistically significant difference was observed in the cell cycles among the three groups(P>0.05).In addition,Western blot showed that the expression of the caspase-3 protein was significantly higher and that of the BCL2 protein markedly lower in the siRNA-SEMG1 than in the blank and negative control groups(P<0.05).Conclusion:SEMG1-specific siRNA can effectively silence the expression of the SEMG1 protein in GC-1 spg cells and promote their apoptosis.