中山大学学报（医学科学版）2011，Vol.32Issue(4)：442-448,510,8.
RNAi干扰WT1和Pax2表达对逆转肾小管上皮间充质转化的作用
Reverse Effect on Renal Tubular Epithelial to Mesenchymal Transition by RNAi Silencing WT1 and Pax2 Expression
摘要
Abstract
[Objective] To explore the reverse effect on renal tubular epithelial to mesenchymal transition by RNA interference (RNAi) silencing the embryonic genes WT1 and Pax2 expression. [Methods] RNAi was used to inhibit WT1 and Pax2. WT1 shRNA expression vector pshRNA-WT1-452 and pshRNA-WTl-283, Pax2 shRNA expression vector pshRNA-Pax2-983 and pshRNA-Pax2-302 were constructed. Plasmids were transfected into NEK52E cells with cationic liposome, and then 10 ng/mL IL-1α was added to stimulate NRK52E cells. RNA and protein were extracted at different time points respectively, and WT1, Pax2, Snail, the epithelial marker E-cadherin, the mesenchymal marker α-SMA mRNA and protein were detected using RT-PCR and Western blot analysis; the morphology of cells was observed. [Result] pshRNA-WTl and pshRNA-Pax2 specifically and efficiently inhibited WT1 and Pax2 expression in NEK52E cells by RNAi, the repression rates were 80.4% and 82.7%, respectively. Repressing WT1 and Pax2 using RNAi blocked EMT in NRK52E cells treated with IL-la, which was evidenced by suppressed a-SMA, Snail expression, restored E-cadherin expression, and the normal cell morphology. [Conclusion] Our experiments suggest that WT1 and Pax2 expression in tubular epithelial cells play an important role in promotion of EMT, and there may be therapeutic value in silencing WT1 and Pax2 so as to prevent or to reverse renal fibrosis.关键词
RNAi；上皮间充质转化；肾小管上皮细胞Key words
RNA interference/epithelial to mesenchymal transition/renal tubular epithelial cells分类
医药卫生引用本文复制引用
黄彬,姜傥,皮蕾,滕军旗..RNAi干扰WT1和Pax2表达对逆转肾小管上皮间充质转化的作用[J].中山大学学报（医学科学版）,2011,32(4):442-448,510,8.基金项目
国家自然科学基金(30370661)；广东省自然科学基金(31683) （30370661）
