Jiang Lina, Jiang Yizhou, Ji Xiaohui, Li Jiangtao, Zhai Ximei
Department of Plastic Surgery, The First Affiliated Hospital of Zhengzhou University No. 1, East Jianshe Rd Zhengzhou 450052 China
Department of Breast Surgery, Shanghai Cancer Center, Fudan University Shanghai 200032 China.
RSC Adv. 2019 Jul 5;9(37):21125-21133. doi: 10.1039/c8ra10552a.
MicroRNAs (miRNAs) are involved in multiple skin pathologies, including wound healing. Here, we explored the detailed role and molecular mechanism of miR-132 on HaCaT cells proliferation and migration. qRT-PCR assay was used to assess miR-132 expression and Western blot analysis was performed to detect inhibitor of matrix metalloproteinase-3 (TIMP3) level in HaCaT cells and normal human epidermal keratinocytes (NHEK) under transforming growth factor β1 (TGF-β1) treatment. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were employed to confirm the endogenous interaction between miR-132 and TIMP3. Cell proliferation ability was determined by MTT assay and the migration capacity was evaluated by transwell assay. TGF-β1 treatment resulted in a increase of miR-132 expression and a decrease of TIMP3 level in HaCaT cells and NHEK cells. The proliferation and migration abilities of TGF-β1-treated HaCaT cells were promoted by miR-132 upregulation, while them were inhibited by TIMP3 overexpression. Moreover, TIMP3 was a direct target of miR-132. MiR-132-mediated pro-proliferation and pro-migration effects were antagonized by TIMP3 in HaCaT cells under TGF-β1 treatment. Our data supported that miR-132 promoted the proliferation and migration of HaCaT cells at least partly by targeting TIMP3, highlighting miR-132 as a potential therapeutic strategy of wound healing.
微小RNA（miRNA）参与多种皮肤病理过程，包括伤口愈合。在此，我们探讨了miR-132对HaCaT细胞增殖和迁移的具体作用及分子机制。采用qRT-PCR检测miR-132表达，通过蛋白质印迹分析检测转化生长因子β1（TGF-β1）处理下HaCaT细胞和正常人表皮角质形成细胞（NHEK）中基质金属蛋白酶-3抑制剂（TIMP3）水平。采用双荧光素酶报告基因检测和RNA免疫沉淀（RIP）检测来证实miR-132与TIMP3之间的内源性相互作用。通过MTT法测定细胞增殖能力，采用Transwell法评估迁移能力。TGF-β1处理导致HaCaT细胞和NHEK细胞中miR-132表达增加，TIMP3水平降低。上调miR-132可促进TGF-β1处理的HaCaT细胞的增殖和迁移能力，而过表达TIMP3则抑制其增殖和迁移能力。此外，TIMP3是miR-132的直接靶点。在TGF-β1处理下，TIMP3可拮抗miR-132介导的HaCaT细胞促增殖和促迁移作用。我们的数据支持miR-132至少部分通过靶向TIMP3促进HaCaT细胞的增殖和迁移，突出了miR-132作为伤口愈合潜在治疗策略的作用。
