MiR-484通过靶向MAPK8抑制人前体脂肪细胞的增殖与分化
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南京医科大学第一附属医院江苏省人民医院儿童保健科

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国家自然科学基金,江苏省妇幼健康科研项目面上项目


MiR-484 inhibits human pre-adipocyte differentiation and proliferation by targeting MAPK8
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Department of Child Health Care, the First Affiliated of Nanjing Medical University

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National Youth Science Fund Project of National Natural Science Foundation of China,Maternal and Child Health Research Project in Jiangsu Province

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    摘要:

    [摘要]:目的:探讨miRNA-484在肥胖发生中的作用及其临床意义。方法:利用RT-qPCR 、CCK-8法、油红O染色及甘油三酯定量检测研究miR-484对人脂肪细胞增殖及成脂分化的作用。运用Targetscan7.2对miR-484的下游靶基因进行预测分析,并使用双荧光素酶实验验证miR-484与靶基因的结合,使用蛋白免疫印迹检测(Western blot)及RT-qPCR验证miR-484对靶基因作用。结果:miR-484在人脂肪细胞分化成熟过程中逐渐低表达。过表达miR-484抑制人前体脂肪细胞的增殖及分化,而沉默miR-484与过表达作用相反。生物信息学预测发现MAPK8是miR-484的下游靶基因,且miR-484可抑制其mRNA及蛋白表达,同时过表达MAPK8可部分挽救miR-484对人前体脂肪细胞增殖分化的抑制。结论:miR-484可以通过抑制MAPK8的表达从而抑制人前体脂肪细胞增殖与成脂分化。

    Abstract:

    [Abstract]: Objective: To investigate the role of miR-484 in adipogenesis and its clinical potentiality. Methods: To explore the function of miR-484 in human pre-adipocyte(HPA) differentiation and proliferation, RT-qPCR, CCK-8, red oil staining and triglyceride content assays were conducted. Additionally, a dual-luciferase reporter assay demonstrated MAPK8, which predicted by Targetscan7.2, was a direct target gene of miR-484 during preadipocyte differentiation, and the results were confirmed by RT-qPCR and Western blot. Results: The results show that miR-484 is down-regulated during HPA differentiation. Compeared with NC, miR-484 inhibits the proliferation and differentiation in HPA, and miR-484 has the opposite effects . A dual-luciferase reporter assay demonstrated MAPK8 is a direct target gene of miR-484, additionally, overexpression of MAPK8 may reserve the miR-484 negative act on pre-adipocyte differentiation and proliferation. Conclusion: MiR-484 inhibits HPA differentiation and proliferation by targeting MAPK8.

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  • 收稿日期:2021-07-26
  • 最后修改日期:2021-11-19
  • 录用日期:2022-03-11
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