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MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

机译:MiR-132通过靶向TIMP3来增强HACAT细胞的增殖和迁移

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摘要

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 beta 1 (TGF-beta 1) 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-beta 1 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-beta 1-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-beta 1 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.
机译:MicroRNAS(miRNA)参与多种皮肤病理,包括伤口愈合。在这里,我们探讨了miR-132对HaCat细胞增殖和迁移的详细作用和分子机制。 QRT-PCR测定用于评估miR-132表达,并进行Western印迹分析,以在转化生长因子β1(TGF- β1)治疗。使用双荧光素酶报告器测定和RNA免疫沉淀(RIP)测定以确认MIR-132和TIMP3之间的内源性相互作用。通过MTT测定法测定细胞增殖能力,通过Transwell测定评估迁移能力。 TGF-β1治疗导致MIR-132表达的增加和HACAT细胞和NHEK细胞中的TIMP3水平降低。通过miR-132上调促进了TGF-β1处理的HaCAT细胞的增殖和迁移能力,而它们被TIMP3过表达抑制。此外,TIMP3是MIR-132的直接靶标。 miR-132介导的促液相传和迁移效应在TGF-β1处理下的HACAT细胞中拮抗。我们的数据支持MIR-132至少部分地通过靶向TIMP3来促进HACAT细胞的增殖和迁移,突出miR-132作为伤口愈合的潜在治疗策略。

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  • 来源
    《RSC Advances》 |2019年第37期|共9页
  • 作者单位

    Zhengzhou Univ Affiliated Hosp 1 Dept Plast Surg 1 East Jianshe Rd Zhengzhou 450052 Henan Peoples R China;

    Fudan Univ Shanghai Canc Ctr Dept Breast Surg Shanghai 200032 Peoples R China;

    Dept Pathol Beijing Peoples R China;

    Peoples Hosp Zhengzhou Dept Breast Surg Zhengzhou 450003 Henan Peoples R China;

    Zhengzhou Univ Affiliated Hosp 1 Dept Plast Surg 1 East Jianshe Rd Zhengzhou 450052 Henan Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
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