Effect of WT1 silencing by siRNA on expression of Wnt/β-catenin and nephrin in mouse podocytes |
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Authors: | ZHOU Jing YUAN Wei-jie XIE Yuan-sheng CHEN Xiang-mei |
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Institution: | 1.Department of Nephrology, Shanghai First Peoples Hospital, Shanghai Jiaotong University, Shanghai 200080, China;2.Institute of Nephrology, Chinese PLA General Hospital, Beijing 100853, China. |
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Abstract: | AIM: To study the effect of WT1 silencing by small interfering RNA (siRNA) on podocyte vitality and expression of Wnt/β-catenin and nephrin in mouse podocytes. METHODS: Conditionally immortalized mouse podocytes were cultured at 33 ℃ in RPMI-1640 medium for proliferation and induced for differentiation at 37 ℃. The podocytes were transfected with WT1 siRNA. The cell vitality was detected by MTT assay. The expression of WT1,Wnt1,β-catenin and nephrin at mRNA and protein levels was determined by real-time qRT-PCR and Western blotting. RESULTS: WT1 siRNA induced the increase in the expression of Wnt1 at mRNA and protein levels, inhibited the phosphorylation of β-catenin, and reduced the cell vitality. Meanwhile, the expression of nephrin at mRNA and protein levels was decreased. CONCLUSION: WT1 siRNA reduces the expression of nephrin in podocytes and the vitality of the cells by activating Wnt/β-catenin signaling pathway. |
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Keywords: | WT1 proteins Podocytes Small interfering RNA Wnt/β-catenin signaling pathway Nephrin |
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