Wang J, Qiu Y, Shi N-W, Zhao J-N, Wang Y-C, Jiang H, Qian H-B
Department of Orthopedics, Jinling Hospital, Nanjing, China.
Eur Rev Med Pharmacol Sci. 2016 Sep;20(18):3748-3759.
The aim of this study was to investigate the molecular mechanism into the keratinocyte migration, which is promoted by Transforming growth factor-β1 (TGF-β1) during wound healing.
In the present study, we investigated the regulation by TGF-β1 on phosphatase and tensin homolog (PTEN) expression, and microRNA-21 (miR-21) level with real-time quantitative PCR or/and Western blotting, and then examined the regulatory role of miR-21 on the PTEN expression and the mesenchymal transition, with real-time quantitative PCR, western blotting and luciferase reporter assay, and the migration of keratinocytic HaCaT cells with scratch assay.
It was demonstrated that miR-21 was upregulated by TGF-β1 treatment in HaCaT cells; and the upregulated miR-21 targeted the 3' UTR of PTEN gene and downregulated the PTEN expression, along with the Smad3/4 upregulation. Moreover, the miR-21 manipulation with miR-21 mimics or miR-21 inhibitor not only upregulated or downregulated the miR-21 level, but also associated with the mesenchymal transition and the migration of HaCaT cells via promoting or downregulating the FSP1 and Collagen I and the E-cadherin, and via upregulating or downregulating the migration of HaCaT cells.
Our results demonstrate that miR-21 mediates the TGF-β1-promoted mesenchymal transition and migration of keratinocytes during skin wound healing via targeting PTEN. This study implies that miR-21 might be an important target to promote the skin wound healing.
本研究旨在探究在伤口愈合过程中由转化生长因子-β1（TGF-β1）促进的角质形成细胞迁移的分子机制。
在本研究中，我们通过实时定量PCR或/和蛋白质印迹法研究了TGF-β1对磷酸酶及张力蛋白同源物（PTEN）表达和微小RNA-21（miR-21）水平的调控，然后通过实时定量PCR、蛋白质印迹法和荧光素酶报告基因检测，研究了miR-21对PTEN表达和间充质转化的调控作用，并通过划痕实验检测了角质形成细胞HaCaT细胞的迁移情况。
结果表明，TGF-β1处理可使HaCaT细胞中的miR-21上调；上调的miR-21靶向PTEN基因的3'UTR并下调PTEN表达，同时Smad3/4上调。此外，用miR-21模拟物或miR-21抑制剂处理miR-21不仅上调或下调了miR-21水平，还通过促进或下调FSP1和胶原蛋白I以及E-钙黏蛋白，以及上调或下调HaCaT细胞的迁移，与HaCaT细胞的间充质转化和迁移相关。
我们的结果表明，miR-21在皮肤伤口愈合过程中通过靶向PTEN介导TGF-β1促进的角质形成细胞间充质转化和迁移。本研究表明，miR-21可能是促进皮肤伤口愈合的重要靶点。
