RNA干扰PLK1基因诱导人前列腺癌细胞PC-3的凋亡研究

RNA interference targeting PLK1 induced human prostatic carcinoma cell line PC-3 apoptosis

目的探讨质粒表达载体介导的RNA干扰抑制PLK1基因表达对人前列腺癌细胞PC-3增殖与凋亡的影响。方法构建靶向人PLK1基因的RNA干扰表达质粒(psh-PLK1),将该干扰质粒转染到人前列腺癌细胞PC-3中,应用RT-PCR技术分析RNA干扰后各组细胞PLK1基因mRNA表达水平的变化,采用MTT法和细胞形态学分析检测PLK1基因表达受到抑制后PC-3细胞的增殖情况的变化,通过流式细胞分析技术以及PI染色技术进行对各组细胞进行细胞凋亡检测。结果RT-PCR结果显示,转染RNA干扰表达质粒(psh-PLK1)24h和48h后,实验组PC-3细胞较对照组的PLK1基因mRNA表达分别降低了74.6%和91.2%;转染了psh-PLK的PC-3细胞与对照组相比生长受到明显抑制(P<0.05),而各对照组之间差异无显著性(P>0.05);流式细胞仪检测发现转染了psh-PLK1的PC-3细胞较对照组细胞产生了更强的细胞凋亡(39.2%),PI荧光染色可见实验组细胞呈现出典型的细胞凋亡形态学变化。结论RNA干扰PLK1基因能够有效地抑制PC-3细胞中PLK1基因的表达,从而抑制PC-3细胞的生长与增殖,并诱导PC-3细胞产生凋亡。

[Objection] To investigated the effects of PLK1 depletion on cell proliferation and apoptosis of PC-3 cells, a human prostatic carcinoma cell line. [Methods] A recombinant plasmid containing shRNA targeting PLK1 （psh-PLK1） was transfected into PC-3 cells. Reverse transcription polymerase chain reaction was used to examine PLK1 gene mRNA expression. Cell proliferation was evaluated by MTT assay and cell morphology analysis. Cell apoptosis was examined by flow cytometry （FCM） and PI staining. [Results] After transfection into PC-3 cells, psh- PLK1 reduced PLK1 mRNA by 74.6% for 24 hours and by 91.2% for 48 hours. Abnormal morphological changes of cells and growth inhibition were observed in psh-PLK1 transfected group. Furthermore, 48 hours aftert transfection, PC-3 cells treatmented with psh-PLK1 showed strong cell apoptosis by FCM and PI staining. [Conclusions] Plasmid-mediated PLK1 RNA interference can effectively down-regulate PLK1 gene expression, inhibit cell proliferation and induce cells apoptosis in PC-3 cells.