PIAS3敲低对体外前列腺癌细胞增殖和凋亡的影响

Effects of PIAS3 knocking down on the proliferation and apoptosis of prostate carcinoma cell in vitro

目的研究PIAS3敲低对人前列腺癌细胞系DU145细胞增殖、细胞周期和凋亡的影响。方法构建PIAS3shRNA表达质粒pSilencer4.1/PIAS3,转染DU145细胞。MTT法检测细胞增殖,流式细胞仪分析细胞周期和凋亡。结果测序证实PIAS3shRNA表达质粒构建成功。转染后DU145细胞PIAS3蛋白表达明显下调。MTT分析显示PIAS3敲低促进细胞增殖,并存在剂量-效应关系。流式细胞术分析显示:PIAS3敲低后S期细胞比例增加,G0/G1期细胞比例减少,凋亡细胞比例减少。结论PIAS3敲低促进体外前列腺癌细胞增殖,抑制其凋亡。

Objective To study the effects of PIAS3 knocking down on the proliferation, cell cycle and apoptosis of human prostate cancer cell line DU145 in vitro. Methods PIAS3 specific short hairpin RNA（shRNA） expressing plasmid was constructed and named pSilencer4. 1/PIAS3. DU145 cells were transfected with pSilencer4. 1/ PIAS3. The proliferation of DU145 cells was analyzed by MTT assay. Cell cycle and apoptosis of DU145 cells were analyzed by flowcytometry. Results PIAS3 shRNA expressing plasmid was succefully constructed and then confirmed by sequencing. Expression of PIAS3 in DU145 was significantly reduced after pSilencer4. 1/PIAS3 transfection. MTT assay showned accelerated proliferation after PIAS3 knocking down, and showned dose-effect curve. Flowcytometry showed cells in S phase increased, cells in G0/G1 decreased and percentage of apoptotic cells decreased after PIAS3 knocking down. Conclusion Knocking down of PIAS3 expression accelerates DU145 cell proliferation and inhibit cell apoptosis in vitro.