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Home > Archive>Volume 32, Issue 9, 2022 >1-9. DOI:10. 3969 / j.issn.1671-7856. 2022. 09. 001
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Role of miR-488-3p targeting regulation of RAP1A in homocysteine-mediated hepatocyte autophagy
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Affiliation:

1. School of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, China. 2. School of Basic Medicine,Ningxia Medical University, Yinchuan 750004. 3. Key Laboratory of Metabolic Cardiovascular Disease Research,National Health Commission (Ningxia Medical University), Yinchuan 750004.4. General Hospital of Ningxia Medical University, Yinchuan 750004

Clc Number:

R-33

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    Abstract:

    Objective To investigate the role of the microRNA miR-488-3p in homocysteine-mediated hepatocyte autophagy through targeted regulation of the RAS oncogene family member RAP1A. Methods Human normal hepatocytes(HL-7702) were routinely cultured in vitro and administered 100 μmol / L Hcy for 24 h. Western blot was used to detect expression levels of autophagy-related proteins LC3, p62 and RAP1A in hepatocytes, whereas qRT-PCR was used to detect the expression of miR-488-3p and RAP1A in these cells. Following transfection of miR-488-3p inhibitor and mimics, qRTPCR and western blot were used to detect the transfection efficiency of miR-488-3p and its effect on LC3 and p62 protein expression. TargetScan was used to predict the correlation between miR-488-3p and RAP1A genes, whereas western blotting was used to detect changes in the expression of RAP1A, a potential downstream target protein of miR-488-3p. Pearson’s correlation coefficient was used to evaluate potential correlations between expression levels of miR-488-3p and autophagy-related proteins in hepatocytes. Results Compared with the control group, expression levels of autophagyrelated proteins LC3, RAP1A and miR-488-3p in the Hcy group were increased (P<0. 01), whereas expression of p62 was significantly decreased (P<0. 01). Following administration of miR-488-3p inhibitor and mimics, expression levels of LC3 and RAP1A proteins in the miR-488-3p inhibitor group were significantly decreased (P<0. 01) and expression of p62 was significantly increased (P<0. 01) compared with the NC-inhibitor group. Compared with the NC-mimics group, expression levels of LC3 and RAP1A proteins in the miR-488-3p mimics group were significantly increased ( P < 0. 01 ), and expression of p62 was significantly decreased (P<0. 01). Further mechanistic studies showed that RAP1A is a downstream target gene of miR-488-3p that is positively regulated by this miRNA. Pearson’ s correlation analysis indicated that the expression level of miR-488-3p was positively correlated with protein expression of LC3 ( r= 0. 9329, P= 0. 0002), but negatively correlated with p62 protein expression ( r= - 0. 8086, P= 0. 0083). Conclusions miR-488-3p is highly expressed in Hcy-mediated hepatocytes and can promote their autophagy by targeting expression of RAP1A.

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History
  • Received:January 17,2022
  • Online: January 16,2023
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