TY - JOUR
T1 - MicroRNA-129-5p modulates epithelial-to-mesenchymal transition by targeting SIP1 and SOX4 during peritoneal dialysis
AU - Xiao, Li
AU - Zhou, Xun
AU - Liu, Fuyou
AU - Hu, Chun
AU - Zhu, Xuejing
AU - Luo, Ying
AU - Wang, Ming
AU - Xu, Xiaoxuan
AU - Yang, Shikun
AU - Kanwar, Yashpal S.
AU - Sun, Lin
N1 - Funding Information:
This work was supported by the Natural Sciences Foundation of China (81100541, 81370832, 81270812, 81470960 and 81300600), the Doctoral Fund of Ministry of Education of China (20110162110012), Furong Scholars Fund from Hunan Province Education Department and the NIH Grant (DK60635).
Publisher Copyright:
© 2015 USCAP, Inc All rights reserved.
PY - 2015/7/27
Y1 - 2015/7/27
N2 - Peritoneal dialysis (PD) is the most readily feasible home-dialysis method for renal replacement therapy. However, repeated use of PD can lead to induction of mesothelial/epithelial-mesenchymal transition (MMT/EMT) and fibrosis, eventually leading to ultrafiltration failure and discontinuation of PD. MicroRNA-129-5p (miR-129-5p) is believed to be a potent downstream inhibitor of TGF-β1 in renal fibrosis, but the effect of miR-129-5p on MMT/EMT relevant to PD is unknown. In this study, as determined by microRNA array analysis and confirmed by northern blot analysis and real-time PCR, we demonstrate that miRNA-129-5p is decreased in mesothelial cells isolated from effluent of patients having PD for more than 6 months extending to several years compared with those who have undergone PD for less than 6 months. The decreased expression of miR-129-5p was accompanied with alterations in EMT-related genes and the expression of respective proteins in vivo. In addition, in in vitro studies we noted that the expression of E-cadherin and claudin-1 were significantly reduced with increased cell migration in HMrSV5, a human peritoneal mesothelial cell line (HPMC), treated with TGF-β1, whereas expression of vimentin, fibronectin and transcription factors SIP1 and SOX4 increased significantly, as assessed by real-time PCR, western blot analysis and immunofluorescence microscopy. Furthermore, alteration in EMT-related genes and proteins were reversed by overexpression of miR-129-5p. No effect was observed in cells treated with miR-negative control. Meanwhile, inhibition of SIP1 and SOX4 with their respective siRNA also could decrease the expression of EMT-related genes and protein levels in HPMCs induced with TGF-β1. Finally, we demonstrate that SIP1 can inhibit the promoter activity of E-cadherin while enhancing the promoter activity of vimentin. We also observed that miR-129-5p could directly target the 3′UTR of SIP1 and SOX4 genes, and repressed their post-transcriptional activities. These data suggest that there is a novel TGF-β1/miR-129-5p/SIP-1 or SOX4 pathway that has a significant role in MMT and fibrosis in the setting of PD.
AB - Peritoneal dialysis (PD) is the most readily feasible home-dialysis method for renal replacement therapy. However, repeated use of PD can lead to induction of mesothelial/epithelial-mesenchymal transition (MMT/EMT) and fibrosis, eventually leading to ultrafiltration failure and discontinuation of PD. MicroRNA-129-5p (miR-129-5p) is believed to be a potent downstream inhibitor of TGF-β1 in renal fibrosis, but the effect of miR-129-5p on MMT/EMT relevant to PD is unknown. In this study, as determined by microRNA array analysis and confirmed by northern blot analysis and real-time PCR, we demonstrate that miRNA-129-5p is decreased in mesothelial cells isolated from effluent of patients having PD for more than 6 months extending to several years compared with those who have undergone PD for less than 6 months. The decreased expression of miR-129-5p was accompanied with alterations in EMT-related genes and the expression of respective proteins in vivo. In addition, in in vitro studies we noted that the expression of E-cadherin and claudin-1 were significantly reduced with increased cell migration in HMrSV5, a human peritoneal mesothelial cell line (HPMC), treated with TGF-β1, whereas expression of vimentin, fibronectin and transcription factors SIP1 and SOX4 increased significantly, as assessed by real-time PCR, western blot analysis and immunofluorescence microscopy. Furthermore, alteration in EMT-related genes and proteins were reversed by overexpression of miR-129-5p. No effect was observed in cells treated with miR-negative control. Meanwhile, inhibition of SIP1 and SOX4 with their respective siRNA also could decrease the expression of EMT-related genes and protein levels in HPMCs induced with TGF-β1. Finally, we demonstrate that SIP1 can inhibit the promoter activity of E-cadherin while enhancing the promoter activity of vimentin. We also observed that miR-129-5p could directly target the 3′UTR of SIP1 and SOX4 genes, and repressed their post-transcriptional activities. These data suggest that there is a novel TGF-β1/miR-129-5p/SIP-1 or SOX4 pathway that has a significant role in MMT and fibrosis in the setting of PD.
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U2 - 10.1038/labinvest.2015.57
DO - 10.1038/labinvest.2015.57
M3 - Article
C2 - 25961171
AN - SCOPUS:84933052852
VL - 95
SP - 817
EP - 832
JO - Laboratory Investigation
JF - Laboratory Investigation
SN - 0023-6837
IS - 7
ER -