Jian-Qing Wang1, Fu Qin2, Liang Zhu3. 1. Department of Nephrology, Second Affiliated Hospital of Zhejiang University, Hangzhou, China. 2. Department of Orthopaedics, First People's Hospital of Yuhang District, Hangzhou, China. 3. Department of Rheumatology, Second Affiliated Hospital of Zhejiang University, Hangzhou, China zhuliangdr@163.com.
Abstract
OBJECTIVE: To explore the role of Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) in autosomal-dominant polycystic kidney disease (ADPKD). METHODS: NHERF1 and β-catenin protein were detected by immunohistochemistry and Western blotting of kidney tissue samples from patients with ADPKD and controls (normal kidney tissue [>5 cm from the foci] collected from patients undergoing unilateral nephrectomy for kidney cancer). NHERF1 and β-catenin protein and mRNA were quantified by Western blot and real-time fluorescent quantitative polymerase chain reaction, respectively, in kidney tissue samples from Han:SPRD (+/+) and (cy/+) rats. The effects of human recombinant NHERF1 on proliferation and cell cycle of ADPKD cyst-lining epithelial cells (WT9-12) were evaluated by MTT assay and flow cytometry, respectively. RESULTS: Levels of NHERF1 protein and mRNA were significantly lower, and β-catenin levels significantly higher, in patients with ADPKD and Han:SPRD (cy/+) rats, compared with control subjects and (+/+) rats, respectively. Exogenous recombinant NHERF1 significantly inhibited proliferation of WT9-12 cells and increased the proportion of cells in G(0)/G(1) phase. CONCLUSIONS: ADPKD is associated with a decrease in NHERF1 protein and mRNA levels. Supplementing exogenous NHERF1 inhibited the proliferation of WT9-12 cells.
OBJECTIVE: To explore the role of Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) in autosomal-dominant polycystic kidney disease (ADPKD). METHODS:NHERF1 and β-catenin protein were detected by immunohistochemistry and Western blotting of kidney tissue samples from patients with ADPKD and controls (normal kidney tissue [>5 cm from the foci] collected from patients undergoing unilateral nephrectomy for kidney cancer). NHERF1 and β-catenin protein and mRNA were quantified by Western blot and real-time fluorescent quantitative polymerase chain reaction, respectively, in kidney tissue samples from Han:SPRD (+/+) and (cy/+) rats. The effects of human recombinant NHERF1 on proliferation and cell cycle of ADPKD cyst-lining epithelial cells (WT9-12) were evaluated by MTT assay and flow cytometry, respectively. RESULTS: Levels of NHERF1 protein and mRNA were significantly lower, and β-catenin levels significantly higher, in patients with ADPKD and Han:SPRD (cy/+) rats, compared with control subjects and (+/+) rats, respectively. Exogenous recombinant NHERF1 significantly inhibited proliferation of WT9-12 cells and increased the proportion of cells in G(0)/G(1) phase. CONCLUSIONS:ADPKD is associated with a decrease in NHERF1 protein and mRNA levels. Supplementing exogenous NHERF1 inhibited the proliferation of WT9-12 cells.