RNAi抑制NEK293细胞TGF-β_1表达的研究  

Inhibitory effect of siRNA on expression of transforming growth factor-β_1 in NEK293 cells

作　　者：崔飞伦[1] 邱镇[2] 陆洪兵[1] 姚震[1] 林大春[1] 

机构地区：[1]江苏大学附属人民医院泌尿外科,江苏镇江212002 [2]解放军第359医院泌尿外科,江苏镇江212000

出　　处：《第四军医大学学报》2008年第12期1125-1127,共3页Journal of the Fourth Military Medical University

摘　　要：目的:探讨针对转化生长因子β1(TGF-β1)的小干扰RNA(siRNA)对大鼠肾间质细胞系(NEK293)TGF-β1的表达、细胞周期的抑制及细胞凋亡的诱导作用.方法:利用RNA干扰(RNAi)效应设计针对TGF-β1的不同siRNA,构建表达载体质粒并转染NEK293细胞系.RT-PCR和WesternBlot分别检测TGF-β1mRNA及蛋白的表达,细胞生长试验和流式细胞观察转染前后细胞周期及细胞凋亡的变化.另设一无意义RNA载体质粒为阴性对照.结果:siRNA转染效率70%,特异性siRNA对TGF-β1mRNA及蛋白的表达具有抑制作用(P>0.05),实验组细胞生长受到抑制、凋亡增加(P>0.05).结论:应用RNAi技术可明显抑制TGF-β1的表达,并高效的抑制NEK293的生长、诱导其凋亡.AIM： To explore the effects of transforming growth factor （TGF）-β1 -specific siRNAs on TGF-β1 expression, cell circle and apoptosis of NEK293 cells. METHODS ： Three TGF-β1 - specific siRNAs were selected with the suppressing effect. RNAi expression vectors were constructed and transfected into NEK293 cells. RT-PCR and Western Blot were used to detect the expression of TGF-β1. The proliferation of the cells was measured by MTT test. The apoptosis of NEH293 cells was measured by flow cytometry. A nonsense small RNA was set as negative control. RESULTS： The greatest transfection efficiency in experimental groups was 70%. When compared with the control groups, the TGF-β1 -specific siRNAs inhibited the expression of TGF-β1 and suppressed the proliferation and induced the apoptosls of the NEK293 cells （P 〈 0.05 ）. CONCLUSION： The TGF-β1-specific siRNAs can inhibit the expression of TGF-β1 , suppress the proliferation, and induce the apoptosis of the NEK293 cells.

关 键 词：转化生长因子Β1 肾纤维化 RNA干扰 

分 类 号：R692[医药卫生—泌尿科学]