Gli1小分子干扰RNA对人胰腺癌细胞增殖和凋亡的影响

Effects of Gli1 siRNA-mediated gene silencing on proliferation and apoptosis of human pancreatic cancer cell

目的:研究锌指转录因子Gli1小分子干扰RNA(siRNA)对人胰腺癌PC-2细胞的增殖和凋亡的影响。方法:筛选出最佳的靶向Gli1siRNA转染人胰腺癌PC-2细胞,采用逆转录-聚合酶链反应(RT-PCR)和Western印迹分别从mRNA和蛋白水平检测抑制效果,四甲基偶氮唑蓝(MTT)和TUNEL+PI双染流式细胞检测Gli1被抑制后细胞增殖和细胞凋亡的情况。结果:Gli1siRNA能在mRNA和蛋白水平下调人胰腺癌细胞株PC-2中Gli1基因的表达,以转染72h时最显著。Gli1表达被抑制后,转染后72h时PC-2细胞生长受到明显抑制,凋亡细胞显著增多。结论:Gli1与胰腺癌细胞的生长和凋亡密切相关,抑制胰腺癌细胞中Gli1表达可抑制胰腺癌细胞生长,促进胰腺癌细胞凋亡。

Objective：To investigate the effect of glioma-associated oncogene homolog 1（GLI1）specific siRNA on the cell proliferation and apoptosis of Gli1 up-regulating human pancreatic cancer cells.Methods：Human pancreatic cancer cells of the line PC-2 were cultured and transfected with the most optimal Gli1 siRNA by screening.RT-PCR and Western blot were used to detect the expression of Gli1 mRNA and protein.MTT method was used to examine the proliferation of the cells.The apoptosis of the cells was detected by PI and FITC double-staining.Results：The expression of Gli1 mRNA and protein in the Gli1 siRNA group decreased remarkably,especially after 72 hours transfection.The proliferation of PC-2 cells transfected with Gli1 siRNA did not changed significantly 24 hours after the transfection,however,decreased 48 hours and especially after 72 hours and 1 week transfection,meanwhile,the apoptosis of the PC-2 cells transfected with Gli1 siRNA was significantly higher than that in the cells transfected with the negative Gli1 siRNA and that of the untransfected cells.Conclusion：Gli1 is closely related to the proliferation and apoptosis of tumor cells,transfection of the specific siRNA targeting on Gli1 helps inhibit the expression of Gli1,thus inhibiting the growth and enhancing the apoptosis of the tumor cells.