靶向沉默核干因子对前列腺癌PC-3细胞增殖能力的影响

Silencing Nucleostemin Expression Reduces the Proliferation of PC-3 Cells

目的:检测前列腺癌PC-3、LNCaP及DU145细胞中核干因子(Nucleostemin,NS)基因的表达,研究NS基因沉默后对PC-3细胞增殖能力的影响。方法:采用免疫细胞化学法及逆转录-聚合酶链反应(RT-PCR)分别检测NS蛋白及mRNA在3种前列腺癌细胞中的表达。用NS特异性小发夹RNA表达质粒转染PC-3细胞,分别用RT-PCR及Western印迹方法检测转染后细胞(简称NS-shRNA-PC-3)中NSmRNA及蛋白的变化。比较NS基因沉默前后PC-3细胞体外、裸鼠体内增殖能力及凋亡情况的变化。结果:3种细胞中均显示NS基因高表达。转染后NS-shRNA-PC-3细胞中NS表达显著降低,细胞增殖速度减慢,G0/G1期细胞百分率显著升高,早期凋亡细胞增多。体内致瘤实验显示,NS基因沉默后,PC-3细胞在裸鼠体内增殖能力显著降低。结论:NS在前列腺癌细胞系中呈高表达,RNA干扰沉默NS基因后PC-3细胞增殖能力显著降低,凋亡细胞增多。

Objective： To detect the expression of the nucleostemin （NS） gene in prostate cancer PC-3, LNCaP and DU145 cells, and to study the effect of the NS gene on the proliferation of PC-3 cells after its silencing. Methods ： The protein and mRNA expressions of NS in PC-3, LNCaP and DU145 cells were respectively detected by immunohistochemical staining and RT-PCR. An NS-specific short-hairpin RNA （shRNA） expression plasmid was used to transfect the PC-3 cells （ NS-shRNA-PC-3 ） , followed by observation of the changes of the NS gene and the proliferation and apoptosis of the cells. Results ： The NS gene was highly expressed in the three types of cells. After the transfeetion, the NS expression and the proliferation of the NS-siRNA-PC-3 cells were remarkably reduced, while the percentage of the GO/Gt cells and the early apoptosis of the PC-3 cells obviously increased. A marked decrease was observed in the neoplasm forming ability of the NS-siRNA-PC-3 cells in the nude mice. Conclusion ： NS is highly expressed in prostate cancer cells. The proliferation of PC-3 cells is remarkably reduced and the early apoptosis of PC-3 cells increased after silencing the NS gene by NS-specific shRNA.