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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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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.
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。使用QRT-PCR测定评估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处理导致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TIMP3过表达抑制它们。此外,TIMP3是MIR-132的直接靶标。 MiR-132介导的促进促进和迁移效应被TGF-β1处理下的HACAT细胞中的TIMP3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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