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MicroRNA-146a-5p Attenuates Fibrosis-related Molecules in Irradiated and TGF-beta1-Treated Human Hepatic Stellate
Bao-Ying Yuan1, Yu-Han Chen1,2, Zhi-Feng Wu1
1Department of Radiation Oncology, Zhongshan Hospital, Fudan University, Shanghai, China.
Abstract:
MicroRNAs (miRNAs) have been shown to play a pivotal role in the pathogenesis and maintenance of liver fibrosis by altering expression of their downstream target genes. However, their role in radiation-induced liver fibrosis has not been assessed in detail. Here, we investigated the role of miR-146a-5p and the target gene in regulation of fibrosis-related markers in the human hepatic stellate cell line LX2. LX2 cells were stimulated with 8 Gy of X rays and various concentrations of TGF-β1 (0-5 ng/ml). Expression of α-SMA, collagen 1 and miR-146a-5p was evaluated. The MiR-146a-5p target gene predictions were performed using bioinformatics analysis and confirmed by dual-luciferase reporter experiment. The effect of miR-146a-5p and the involved target gene on the expression of these fibrogenic molecules was also assessed. Expression of α-SMA and collagen 1 were upregulated in response to radiation and/or TGF-β1 treatment and miR-146a-5p levels were altered in LX2 cells. Restoration of miR-146a-5p expression suppressed expression of α-SMA and collagen 1 in irradiated and TGF-β1-treated LX2 cells. Subsequent mechanism experiments revealed that miR-146a-5p overexpression inhibited PTPRA expression by binding to its 3'-untrans-lated region and reduced SRC activation. In addition, enhancement of PTPRA partially reversed the suppressive effect of miR-146a-5p on α-SMA and collagen 1 expression in LX2 cells. In conclusion, miR-146a-5p may negatively regulate the PTPRA-SRC signaling to inhibit expression of fibrosis-related markers in irradiated and TGF-β1-stimulated LX2 cells.
Insights
MicroRNA-146a-5p (miR-146a-5p) can reduce liver fibrosis markers like alpha-SMA and collagen 1. It functions by inhibiting PTPRA and SRC signaling in radiation- or TGF-β1-stimulated liver cells.
Area of Science:
- Molecular Biology
- Cell Biology
- Oncology
Background:
- MicroRNAs (miRNAs) are key regulators in liver fibrosis pathogenesis by modulating target gene expression.
- The specific role of miRNAs in radiation-induced liver fibrosis remains underexplored.
- Human hepatic stellate cells (LX2) are crucial in liver fibrogenesis.
Purpose of the Study:
- To investigate the role of miR-146a-5p in radiation-induced liver fibrosis.
- To identify miR-146a-5p target genes involved in regulating fibrosis markers.
- To elucidate the mechanism by which miR-146a-5p affects fibrogenesis in LX2 cells.
Main Methods:
- LX2 cells were exposed to 8 Gy X-rays and varying TGF-β1 concentrations.
- Expression of alpha-smooth muscle actin (α-SMA), collagen 1, and miR-146a-5p was quantified.
- Bioinformatics and dual-luciferase reporter assays identified miR-146a-5p targets; PTPRA was confirmed.
Main Results:
- Radiation and/or TGF-β1 upregulated α-SMA and collagen 1, altering miR-146a-5p levels in LX2 cells.
- Restoring miR-146a-5p suppressed α-SMA and collagen 1 expression in treated LX2 cells.
- miR-146a-5p inhibited PTPRA expression, reducing SRC activation; PTPRA enhancement reversed miR-146a-5p's suppressive effects.
Conclusions:
- miR-146a-5p negatively regulates PTPRA-SRC signaling, thereby inhibiting fibrosis markers in irradiated and TGF-β1-stimulated LX2 cells.
- This study highlights miR-146a-5p as a potential therapeutic target for radiation-induced liver fibrosis.
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