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LOC102553417 silencing facilitates the apoptosis of hepatic stellate cells via the miR‑30e/MTDH axis.

ABSTRACT: Hepatic fibrosis is an inevitable pathological process in the progression of multiple chronic liver diseases and remains a major challenge in the treatment of liver diseases. The purpose of the present study was to demonstrate whether silencing of the long non‑coding RNA LOC102553417 promoted hepatic stellate cell (HSC) apoptosis via the microRNA (miR)‑30e/metadherin (MTDH) axis. A LOC102553417 silencing lentivirus was constructed and transduced into HSC‑T6 cells. After confirming the silencing efficiency by reverse transcription‑quantitative PCR, cell proliferation was assessed using the Cell Counting Kit‑8 assay and apoptosis was assessed using flow cytometry. The interaction between LOC102553417 and miR‑30e, and that between miR‑30e and MTDH, was demonstrated using the dual‑luciferase reporter assay and RNA binding protein immunoprecipitation. The apoptosis of HSC‑T6 cells was detected after transfection of miR‑30e mimics and inhibitors with or without silencing LOC102553417. Silencing of LOC102553417 curbed HSC‑T6 cell proliferation and expedited their apoptosis. LOC102553417 was demonstrated to target miR‑30e, whereas miR‑30e targeted MTDH. In addition, LOC102553417 silencing significantly upregulated miR‑30e expression levels, and significantly downregulated MTDH mRNA and protein expression levels, which resulted in a significantly reduced p‑Akt/Akt ratio and significantly elevated p53 protein expression levels. Transfection with miR‑30e mimic alone significantly enhanced HSC‑T6 cell apoptosis and inhibits LOC102553417 and MTDH expressions, In addition, miR‑30e mimic expedites the apoptosis of HSCs stimulated by LOC102553417 silencing; consistent results were obtained by reverse validation of miR‑30e inhibitor. In conclusion, the present study demonstrated that LOC102553417 silencing stimulated the apoptosis of HSCs via the miR‑30e/MTDH axis.

SUBMITTER: Wei W

PROVIDER: S-EPMC9551410 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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LOC102553417 silencing facilitates the apoptosis of hepatic stellate cells via the miR‑30e/MTDH axis.

Wei Wujun W Lin Cheng C Hu Rentong R Huang Jingjing J Chen Xiaohao X Zhou Lv L Wei Jiazhu J Deng Yi-Bin YB Wang Chun-Fang CF

Molecular medicine reports 20220930 5

Hepatic fibrosis is an inevitable pathological process in the progression of multiple chronic liver diseases and remains a major challenge in the treatment of liver diseases. The purpose of the present study was to demonstrate whether silencing of the long non‑coding RNA LOC102553417 promoted hepatic stellate cell (HSC) apoptosis via the microRNA (miR)‑30e/metadherin (MTDH) axis. A LOC102553417 silencing lentivirus was constructed and transduced into HSC‑T6 cells. After confirming the silencing ...[more]

PMID: 36177898

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Project description:Background:Microvascular obstruction (MVO) can result in coronary microcirculation embolism and myocardial microinfarction. Myocardial injury induced by MVO is characterized by continuous ischemia and hypoxia of cardiomyocytes. Autophagy and apoptosis are closely associated with various cardiovascular diseases. Based on our previous study, we observed a decrease in miR-30e-3p expression and an increase in Egr-1 expression in a rat coronary microembolization model. However, the specific function of miR-30e-3p in regulating autophagy and apoptosis in an ischemia/hypoxia (IH) environment remains to be deciphered. We exposed cardiomyocytes to an IH environment and then determined whether miR-30e-3p was involved in promoting cardiomyocyte autophagy and inhibiting apoptosis by regulating Egr-1. Methods:Cardiomyocytes were isolated from rats for our in vitro study. miR-30e-3p was either overexpressed or inhibited by transfection with lentiviral vectors into cardiomyocytes. 3-Methyladenine (3-MA) was used to inhibit autophagy. RT-qPCR and western blotting were used to determine the expression levels of miR-30e-3p, Egr-1, and proteins related to the autophagy and apoptosis process. Autophagic vacuoles and autophagic flux were evaluated using transmission electron microscopy (TEM) and confocal microscopy, respectively. Cardiomyocyte viability was evaluated using the MTS assay. Cell injury was assessed by lactate dehydrogenase (LDH) leakage, and apoptosis was determined by flow cytometry. Results:Both miR-30e-3p expression and autophagy were significantly inhibited, and apoptosis was increased in cardiomyocytes after 9 hours of IH exposure. Overexpression of miR-30e-3p increased autophagy and inhibited apoptosis, as well as suppressed Egr-1 expression and decreased cell injury. In addition, inhibition of miR-30e-3p reduced autophagy and increased apoptosis and cell injury. Conclusions:miR-30e-3p may be involved in promoting cardiomyocyte autophagy and inhibiting apoptosis by indirectly regulating Egr-1 expression in an IH environment.

Dataset Information

LOC102553417 silencing facilitates the apoptosis of hepatic stellate cells via the miR‑30e/MTDH axis.

Publications

LOC102553417 silencing facilitates the apoptosis of hepatic stellate cells via the miR‑30e/MTDH axis.

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