MicroRNA-146a-5p Attenuates Fibrosis-related Molecules in Irradiated and TGF-beta1-Treated Human Hepatic Stellate Cells by Regulating PTPRA-SRC Signaling
MicroRNA-146a-5p Attenuates Fibrosis-related Molecules in Irradiated and TGF-beta1-Treated Human Hepatic Stellate Cells by Regulating PTPRA-SRC Signaling
复制标题
MicroRNA-146a-5p 通过调节 PTPRA-SRC 信号传导减弱经辐射和 TGF-β1 处理的人肝星状细胞中的纤维化相关分子
DOI：
10.1667/rr15401.1
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发表时间：
2019-12-01
影响因子：
3.4
通讯作者：
Zeng, Zhao-chong
中科院分区：
文献类型：
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作者：
Yuan, Bao-ying;Chen, Yu-han;Zeng, Zhao-chong
MicroRNA（miRNAs）通过改变其下游靶基因的表达，在肝纤维化的发生和维持中起着关键作用。然而，它们在辐射诱导的肝纤维化中的作用尚未得到详细评估。在此，我们研究了miR-146 a-5 p和靶基因在人肝星状细胞系LX 2中调节纤维化相关标志物的作用。用8戈伊X射线和不同浓度的TGF-β1（0-5 ng/ml）刺激LX 2细胞。评价α-SMA、胶原1和miR-146 a-5 p的表达。MiR-146 a-5 p靶基因的预测使用生物信息学分析进行，并通过双荧光素酶报告基因实验证实。还评估了miR-146 a-5 p和所涉及的靶基因对这些纤维化分子表达的影响。α-SMA和胶原1的表达在辐射和/或TGF-β1处理的响应中上调，并且在LX 2细胞中miR-146 a-5 p水平改变。miR-146 a-5 p表达的恢复抑制了照射和TGF-β1处理的LX 2细胞中α-SMA和胶原1的表达。随后的机制实验表明，miR-146 a-5 p过表达通过与PTPRA的3′-非翻译区结合抑制其表达，降低SRC的激活。此外，PTPRA的增强部分逆转了miR-146 a-5 p对LX 2细胞中α-SMA和胶原1表达的抑制作用。结论：miR-146 a-5 p可能通过负调控PTPRA-SRC信号通路，抑制照射和TGF-β1刺激的LX 2细胞中纤维化相关标志物的表达。
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.