siRNA抑制人前列腺癌PC-3细胞株VEGF的表达及细胞增殖

Inhibition of siRNA on VEGF expression in PC-3 cell line and cell proliferation

目的:研究小干扰RNA(siRNA)对PC-3人前列腺癌细胞血管内皮生长因子(VEGF)基因表达的抑制作用及抑制癌细胞增殖和诱导凋亡的作用。方法:体外合成特异性靶向人VEGF基因的siRNA,并转染到PC-3细胞中;RT-PCR和Western blotting检测siVEGF对VEGF基因和蛋白表达的抑制作用;MTT法检测siVEGF对癌细胞增殖抑制作用,流式细胞术检测siVEGF诱导细胞凋亡的作用。结果:转染siVEGF1和siVEGF2组VEGF mRNA表达水平与空白对照组比较分别下调了60.32%和70.73%;VEGF蛋白表达水平与空白对照组比较明显受到抑制,最高抑制率达72%;转染阴性对照质粒组VEGF基因和蛋白表达水平都无明显改变。转染siVEGF1和siVEGF2的细胞增殖受到抑制,增殖抑制率分别为49.5%和55.3%,细胞的凋亡率分别为33.9%和42.0%。结论:构建的siRNA VEGF能特异有效下调VEGF基因的表达,瞬时转染siRNA VEGF能有效抑制人前列腺癌细胞PC-3增殖并诱导其凋亡。

Objective To study the inhibitory effect of small interfering RNA （siRNA） on VEGF expression in prostate cancer cell line PC-3 and to observe its influences in cell proliferation and apoptosis. Methods Two siRNAs against the VEGF gene were designed and transfected into PC-3 cells respectively, VEGF gene and protein expressions were measured by RT-PCR and Western blotting. Cell proliferation was measured with MTT assay, apoptosis was measured with flow cytometry （FCM）. Results The VEGF mRNA expressions were down- regulated by 60.32 % and 70.73%, and the VEGF protein expressions were maximally down-regulated.by 72% after transfecting with siVEGF1 and siVEGF2, respectively. The inhibitory rates of cell proliferation were 49.5% and 55.3 % the apoptotic rates after transfecting were 33.9 % and 42.0%. Conclusion siRNA against VEGF could effectively down-regulate VEGF expression in PC-3 cell line, it can inhibit cell growth and induce apoptosis.