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RUNX1-mediated repression of miR-24 promotes hepatic stellate cell activation and liver fibrosis by targeting the
Zhijing Wang1, Meiyi Song1, Yingying Zhao2
1Department of Gastroenterology and Hepatology, Tongji Hospital, School of Medicine, Tongji University, Shanghai, China.
Background:
Epigenetic mechanisms, including microRNAs (miRNAs), are increasingly recognized as crucial regulators of organ fibrosis. In this study, we investigated the role of miR-24 in hepatic stellate cell (HSC) activation and liver fibrosis.
Methods:
miR-24 expression was analyzed in carbon tetrachloride (CCl4)-induced liver fibrosis and activated HSCs using quantitative real-time PCR (qRT-PCR). Gain- and loss-of-function experiments of miR-24 were performed in vitro. Western blotting, qRT-PCR, 5-ethynyl-2'-deoxyuridine (EdU) staining, flow cytometry, luciferase reporter assays, bioinformatics analysis, and chromatin immunoprecipitation PCR (ChIP-PCR) were performed to examine the molecular mechanisms of miR-24. Serum miR-24 levels were measured in patients with liver cirrhosis and further analyzed by subgroup.
Results:
We observed significant downregulation of miR-24 in CCl4-induced liver fibrosis and activated HSCs. Functional assays showed that miR-24 overexpression markedly inhibited HSC activation and migration, whereas miR-24 inhibition had the opposite effects. Mechanistically, ALK4 was identified as a direct target of miR-24: miR-24 bound the 3'UTR of ALK4 mRNA, thereby suppressing Smad3 phosphorylation and downstream fibrosis-associated signaling pathways. Furthermore, the transcription factor RUNX1 was induced during HSC activation, and it transcriptionally repressed miR-24 expression. Clinically, serum miR-24 levels were significantly lower in patients with liver cirrhosis than in healthy controls and were negatively correlated with Child-Pugh grade.
Conclusion:
Our findings suggest that the RUNX1/miR-24/ALK4 axis plays a crucial role in HSC activation and migration. miR-24 may serve as a biomarker for liver fibrosis screening, representing a potential therapeutic target for anti-fibrotic intervention.
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