基质交联分子1对前列腺癌PC-3细胞凋亡相关蛋白表达的影响

Inhibition of stromal interaction molecule 1 and the expression of apoptosis-related proteins in prostate cancer PC-3 cells

目的:观察抑制基质交联分子1(STIM1)对前列腺癌PC-3细胞凋亡相关蛋白表达的影响。方法:将携带STIM1基因的小干扰RNA(shRNA)慢病毒载体STIM1-pGCSIL-GFP转染人激素非依赖性前列腺癌PC-3细胞,3d后荧光倒置显微镜观察转染效率;1周后RT-PCR及Western印迹验证STIM1抑制表达有效性,并采用Western印迹检测PC-3细胞中凋亡相关蛋白Bcl-2、Bax,survivin、激活型Caspase-3的表达水平。结果:倒置显微镜观察发现PC-3细胞病毒转染效率>80%。转染1周后,RT-PCR及Western印迹显示STIM1被有效抑制。抑制STIM1表达后,对照组Bcl-2/Bax比率为1.24,干扰组PC-3细胞Bcl-2/Bax比率为0.31,比率显著下降;干扰组PC-3细胞survivin表达明显降低,相对表达量为对照组的0.14倍;Caspase-3裂解激活相对表达量为对照组的1.52倍(P<0.05)。结论:STIM1在前列腺癌PC-3细胞中可视为致癌基因,抑制其表达可通过下调Bcl-2/Bax比率,降低survivin表达,激活Caspase-3级联通路,诱导细胞凋亡。

Objective ： To explore the effects of stromal interaction molecule 1 （STIM1） on the expression of apoptosis-related proteins in prostate cancer PC-3 cells. Methods ： We transfected the lentivirus vector STIMI-pGCSIL-GFP carrying ST1M1 shRNA in- to human hormone-independent prostate cancer PC-3 cells, and 3 days later observed the transfection efficiency by fluorescence micros- copy. At 7 days after transfection, we determined the expression of STIM1 in the PC-3 cells by RT-PCR and Western blot and those of apoptosis-related proteins Bcl-2, Bax, survivin and activated Caspase-3 by Western blot. Results ： At 3 days, inverted microscopy re- vealed a transfection efficiency of 〉 80%. At 7 days, the STIM1 expression was significantly inhibited at both mRNA and protein lev- els. The Bcl-2/Bax rate was remarkably decreased as compared with that of the control group （0.31 vs 1.24 ）, and the survivin ex- pression was markedly reduced, 0.14 times that of the relative expression in the control. However, the Caspase-3 cleavage was signifi- cantly activated, 1.52 times that of the control （ P 〈 O. 05 ）. Conclusion ： STIM1 can be regarded as an oncogene in prostate cancer PC-3 cells. Inhibition of its expression can induce PC-3 cell apoptosis by reducing the Bcl-2/Bax rate, decreasing the survivin expres- sion, and activating the Caspase-3 pathway. Natl J Androl , 2014, 20 （3） ： 225 -228