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微小RNA-185通过靶向过氧化物酶体增殖物激活受体β抑制HaCaT角质形成细胞的增殖和迁移。

MicroRNA-185 inhibits the proliferation and migration of HaCaT keratinocytes by targeting peroxisome proliferator-activated receptor β.

作者信息

Yang Jingzhe, Deng Pingyang, Qi Yonggang, Feng Xinshu, Wen Hailing, Chen Fengping

机构信息

Department of Burn and Plastic Surgery, Affiliated Hospital of Chengde Medical University, South Wing Hospital of Chengde Medical College, Chengde, Hebei 067000, P.R. China.

Department of Burn and Plastic Surgery, Bayingol Mongolia Autonomous Prefecture People's Hospital, Urumqi, Xinjiang Uyghur Autonomous Region 841000, P.R. China.

出版信息

Exp Ther Med. 2021 Apr;21(4):366. doi: 10.3892/etm.2021.9797. Epub 2021 Feb 18.

Abstract

Proliferation and migration of keratinocytes are major processes of skin wound repair after injury. It has been indicated that microRNAs (miRNAs/miRs) are associated with the proliferation and migration of keratinocytes. However, the mechanism by which miR-185 affects these processes in keratinocytes remains unclear. In the present study, the expression level of miR-185 and peroxisome proliferator-activated receptor β (PPARβ) was examined by reverse transcription-quantitative PCR in HaCaT keratinocytes. Cell proliferation was evaluated using Cell Counting Kit-8 and colony formation assays. Western blot analysis was used to detect the levels of cell proliferation, migration and PI3K/AKT signaling pathway-associated proteins. In addition, the migratory capacity of the cells was determined using Transwell assay. The target gene of miR-185 was verified using dual-luciferase reporter assay. The results indicated that overexpression of miR-185 inhibited proliferation, migration and activation of the PI3K/AKT signaling pathway in HaCaT keratinocytes. PPARβ was indicated to be a target of miR-185 and its overexpression promoted the proliferation and migration of HaCaT keratinocytes, while its knockdown exhibited the adverse effects. Furthermore, PI3K inhibitor LY294002 inhibited activation of the PI3K/AKT signaling pathway and decreased the proliferation and migration of HaCaT keratinocytes. In addition, overexpressed PPARβ reversed the suppressive effects of miR-185 overexpression on proliferation, migration and activation of the PI3K/AKT signaling pathway. In conclusion, the results of the present study demonstrated that miR-185 suppressed activation of the PI3K/AKT signaling pathway via targeting PPARβ, thereby regulating proliferation and migration in HaCaT keratinocytes. The present study provided a novel theoretical basis for the use of miR-185 as a target in wound repair.

摘要

角质形成细胞的增殖和迁移是皮肤损伤后伤口修复的主要过程。已有研究表明,微小RNA(miRNA/miR)与角质形成细胞的增殖和迁移有关。然而,miR-185影响角质形成细胞这些过程的机制仍不清楚。在本研究中,通过逆转录定量PCR检测了HaCaT角质形成细胞中miR-185和过氧化物酶体增殖物激活受体β(PPARβ)的表达水平。使用细胞计数试剂盒-8和集落形成试验评估细胞增殖。蛋白质印迹分析用于检测细胞增殖、迁移和PI3K/AKT信号通路相关蛋白的水平。此外,使用Transwell试验测定细胞的迁移能力。使用双荧光素酶报告基因试验验证miR-185的靶基因。结果表明,miR-185的过表达抑制了HaCaT角质形成细胞中PI3K/AKT信号通路的增殖、迁移和激活。PPARβ被证明是miR-185的靶标,其过表达促进了HaCaT角质形成细胞的增殖和迁移,而其敲低则表现出相反的作用。此外,PI3K抑制剂LY294002抑制了PI3K/AKT信号通路的激活,并降低了HaCaT角质形成细胞的增殖和迁移。此外,过表达的PPARβ逆转了miR-185过表达对PI3K/AKT信号通路增殖、迁移和激活的抑制作用。总之,本研究结果表明,miR-185通过靶向PPARβ抑制PI3K/AKT信号通路的激活,从而调节HaCaT角质形成细胞的增殖和迁移。本研究为将miR-185作为伤口修复的靶点提供了新的理论依据。

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