MicroRNA-129-5p modulates epithelial-to-mesenchymal transition by targeting SIP1 and SOX4 during peritoneal dialysis.
MicroRNA-129-5p modulates epithelial-to-mesenchymal transition by targeting SIP1 and SOX4 during peritoneal dialysis.
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MicroRNA-129-5p 在腹膜透析过程中通过靶向 SIP1 和 SOX4 调节上皮间质转化
DOI：
10.1038/labinvest.2015.57
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发表时间：
2015-07
期刊：
影响因子：
--
通讯作者：
Sun L
中科院分区：
文献类型：
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作者：
Xiao L;Zhou X;Liu F;Hu C;Zhu X;Luo Y;Wang M;Xu X;Yang S;Kanwar YS;Sun L
腹膜透析（PD）是肾脏替代治疗中最可行的家庭透析方法。然而，重复使用PD可导致诱导间皮/上皮-间质转化（MMT/EMT）和纤维化，最终导致超滤失败和PD停药。MicroRNA-129- 5 p（miR-129- 5 p）被认为是肾纤维化中TGF-β1的有效下游抑制剂，但miR-129- 5 p对与PD相关的MMT/EMT的影响尚不清楚。在这项研究中，通过microRNA阵列分析确定，并通过北方印迹分析和实时PCR证实，我们证明，与那些经历PD少于6个月的患者相比，从PD超过6个月延伸到数年的患者的流出物中分离的间皮瘤细胞中miRNA-129- 5 p减少。miR-129- 5 p的表达降低伴随着EMT相关基因和相应蛋白在体内的表达的改变。此外，在体外研究中，我们注意到，在用TGF-β1处理的人腹膜间皮细胞系（HPMC）HMrSV 5中，E-cadherin和claudin-1的表达随着细胞迁移的增加而显著降低，而波形蛋白、纤连蛋白和转录因子SIP 1和SOX 4的表达显著增加，如通过实时PCR、蛋白质印迹分析和免疫荧光显微镜所评估的。此外，EMT相关基因和蛋白的改变可通过miR-129- 5 p的过表达逆转。在用miR阴性对照处理的细胞中未观察到作用。同时，siRNA抑制SIP 1和SOX 4也能降低TGF-β1诱导的人乳头状瘤细胞EMT相关基因和蛋白的表达。最后，我们证明SIP 1可以抑制E-钙粘蛋白的启动子活性，同时增强波形蛋白的启动子活性。我们还观察到miR-129- 5 p可以直接靶向SIP 1和SOX 4基因的3′UTR，抑制其转录后活性。这些数据表明，存在一种新的TGF-β1/miR-129- 5 p/SIP-1或SOX 4通路，其在PD背景下的MMT和纤维化中具有显著作用。
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.