RNAi沉默PSA基因对人前列腺癌细胞系LNCaP生长的影响

摘要

Objective To investigate the change of the growth of human prostate cancer cell line LNCaP by silencing PSA genes using RNA interference. Methods Small interfering RNA for PSA gene (PSA-siRNA) was initially constructed. The LNCaP cells were divided into four groups, including PSA-siRNA transfection group, negative control group, transfection reagents group and the control group. Then the expression of PSA was determined by western blot. The apoptosis and proliferation of LNCaP cells were detected by MTT assay and flow cytom-etry. Results After treatment with PSA-siRNA for 48 h, the expression of PSA was significantly decreased compared with the negative control group, transfection reagents group and the control group (P<0.05). PSA-siRNA transfection group has the inhibition rate of 27.5% and 19.3% of cell apoptosis. Conclusion Silencing the PSA gene by RNAi can actively suppress the expression of PSA gene, and inhibit the growth and proliferation of LNCaP cells.%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48 h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.