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目的研究运用RNA干扰技术沉默PSA基因后对人前列腺癌细胞系LNCaP细胞产生的影响。方法构建针对PSA基因的小分子干扰RNA(PSA-siRNA),将前列腺癌LNCaP系进行分组,设PSA-siRNA转染组、阴性对照组、转染液组(只由转染试剂进行转染实验)和对照组(不添加任何转染试剂)进行实验。应用蛋白印迹法检测PSA的变化,并通过MTT法和流式细胞技术检测LNCaP细胞的调亡和细胞增殖方面的变化。结果蛋白印迹法检测结果显示,与阴性对照组、转染液和对照组比较,应用PSA-siRNA处理48h后的LNCaP细胞株,PSA的表达明显降低,组间比较差异有统计学意义(P<0.05)。MTT法和流式细胞仪检测示,转染组的细胞生长抑制率为27.5%,且观察到更强的细胞调亡(19.3%)。结论运用RNA干扰技术沉默PSA基因可以有效抑制LNCaP细胞中PSA基因的表达,进而抑制LNCaP细胞的生长与增殖,并可有效诱导细胞调亡。
Objective To study the effect of RNA interference on silencing PSA gene in human prostate cancer cell line LNCaP. Methods PSA-siRNA was constructed and the prostate cancer LNCaP lines were divided into groups. PSA-siRNA transfection group, negative control group and transfectant group were constructed. Transfection reagent ) And control group (without any transfection reagent). The changes of PSA were detected by Western blotting, and the changes of apoptosis and cell proliferation of LNCaP cells were detected by MTT assay and flow cytometry. Results The results of Western blotting showed that the expression of PSA in LNCaP cells treated with PSA-siRNA for 48 h was significantly lower than that in the negative control group, transfection solution and control group, with significant difference between the two groups (P < 0.05). MTT assay and flow cytometry showed that the transfected group had a cell growth inhibition rate of 27.5% and a stronger apoptosis (19.3%) was observed. Conclusion The silencing of PSA gene by RNA interference can effectively inhibit the expression of PSA gene in LNCaP cells, and then inhibit the growth and proliferation of LNCaP cells and induce cell apoptosis effectively.