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[目的]探讨整合素连接激酶(integrin-linked kinase,ILK)的表达抑制对前列腺癌PC3细胞凋亡的影响。[方法]设计并合成ILK基因的siRNA,同时设立空白对照和阴性对照和阳性实验组,利用脂质体Translipid转染PC3细胞,用半定量RT-PCR和Western-blot检测转染前后PC3细胞ILK mRNA和蛋白的表达水平变化,吖啶橙和嗅乙啶(AO/EB)双重染色检测诱导PC3细胞凋亡的作用,Cell Counting Kit-8(CCK-8)法检测PC3细胞体外增殖的变化。[结果]PC3细胞转染ILKsiRNA 48h后,与空白对照和阴性对照相比,ILK的mRNA表达水平和蛋白表达水平明显下调(31.6%±3.13%、25.3%±2.36%,P<0.01);并诱导34.8%±2.20%(P<0.01)的细胞凋亡;细胞重新种板24、48、72h,与空白对照和阴性对照相比,可以明显抑制细胞增殖,其增殖抑制率分别为9.9%±0.55%、27.8%±1.67%、43.7%±2.0%,P<0.01。[结论]以ILK为靶向的siRNA能够有效下调ILK基因的mRNA和蛋白表达水平,显著抑制PC3细胞的增殖,并能在一定程度上诱导其凋亡。
[Objective] To investigate the effect of inhibition of integrin-linked kinase (ILK) expression on PC3 cell apoptosis in prostate cancer. [Method] siRNA of ILK gene was designed and synthesized. At the same time, blank control, negative control and positive experimental group were set up. PC3 cells were transfected with lipofectamine Translipid. The expression of ILK in PC3 cells was detected by semi-quantitative RT-PCR and Western- The changes of mRNA and protein expression level were detected by acridine orange staining and AO / EB staining. Cell Counting Kit-8 (CCK-8) was used to detect the proliferation of PC3 cells. [Results] Compared with the blank control and the negative control, the mRNA and protein expressions of ILK in PC3 cells transfected with ILK siRNA for 48 h were significantly decreased (31.6% ± 3.13%, 25.3% ± 2.36%, P <0.01). Induced cell apoptosis 34.8% ± 2.20% (P <0.01). The cells were replated 24, 48 and 72 hours later. Compared with the blank control and the negative control, the cell proliferation was inhibited obviously, and the proliferation inhibition rate was 9.9% ± 0.55%, 27.8% ± 1.67%, 43.7% ± 2.0%, P <0.01. [Conclusion] siRNA targeting ILK can effectively down-regulate the mRNA and protein expression of ILK gene, significantly inhibit the proliferation of PC3 cells and induce apoptosis to a certain extent.