文献类型：期刊文献

中文题名：基于miRNA-mRNA调控网络探讨miR-34a-5p靶向Hh通路对LX2细胞增殖的影响

英文题名：Effect of miR-34a-5p targeting the Hh signaling pathway on the proliferation of LX2 cells

作者：任真[1];马燕花[2];王莉[2];王爱娣[2];赵秀萍[2]

第一作者：任真

机构：[1]甘肃中医药大学信息工程学院信息工程教研室,兰州730000;[2]甘肃中医药大学第一临床医学院内科学教研室,兰州730000

第一机构：甘肃中医药大学信息工程学院（教育技术中心）

年份：2023

卷号：52

期号：7

起止页码：582

中文期刊名：中国医科大学学报

外文期刊名：Journal of China Medical University

收录：CSTPCD;;北大核心:【北大核心2020】；

基金：国家自然科学基金(81860821)。

语种：中文

中文关键词：miRNA-mRNA调控网络;微RNA-34a-5p;刺猬信号通路;肝星状细胞;肝纤维化

外文关键词：miRNA-mRNA regulatory network;miR-34a-5p;hedgehog signaling pathway;hepatic stellate cells;liver fibrosis

摘要：目的探讨miR-34a-5p靶向刺猬(Hh)信号通路对肝星状细胞(HSC)增殖和凋亡的调控作用。方法采用生物信息学分析筛选出LX2细胞中靶向Hh信号通路的差异miRNA,通过Cytoscape构建miRNA-m RNA网络。体外培养LX2细胞,通过转化生长因子-β1(TGF-β1)诱导建立LX2细胞活化模型。细胞活力检测试剂盒(CCK-8)检测miR-34a-5p过表达对LX2细胞增值活力的影响;流式细胞分析法(Annexin-V-FITC/PI)检测miR-34a-5p过表达对LX2细胞凋亡的影响;实时荧光定量PCR和Western blotting分析miR-34a-5p过表达对声波刺猬(Shh)、脑胶质瘤相关癌基因1(Gli1)、脑胶质瘤相关癌基因2(Gli2)、Ⅰ型胶原蛋白(Col-Ⅰ)及α平滑肌肌动蛋白(α-SMA)mRNA和蛋白表达水平的影响。结果LX2细胞经TGF-β1诱导活化后,细胞活力显著升高,Shh、Gli1、Gli2、Col-Ⅰ及α-SMA表达均增高(P<0.01)。Hh信号通路特异性阻断剂环靶明脂可降低LX2细胞活力,下调Shh、Gli1、Gli2、Col-Ⅰ及α-SMA蛋白表达(P<0.01)。miR-34a-5p过表达可显著提高LX2细胞活力,上调Gli1、Gli2、Col-Ⅰ及α-SMA蛋白表达(P<0.05)。结论miR-34a-5p靶向调控LX2细胞的Hh信号通路,过表达miR-34a-5p可以上调Hh信号通路相关蛋白表达,促进LX2细胞活化。
Objective This study investigates the regulatory effect of microRNA-34a-5p(miR-34a-5p)on the proliferation and apoptosis of hepatic stellate cells(HSCs)by targeting the hedgehog(Hh)signaling pathway.Methods The differential miRNAs targeting the Hh signaling pathway in LX2 cells were screened using bioinformatics analysis,and the miRNA-mRNA network was constructed using Cytoscape.LX2 cells were cultured in vitro and induced using transforming growth factor-β1(TGF-β1)to establish a LX2 cell activation model.The cell viability assay kit(CCK-8)was used to detect the effect of miR-34a-5p overexpression on the proliferation of LX2 cells.Flow cytometry(Annexin-V-FITC/PI)was used to detect the effect of miR-34a-5p overexpression on the apoptosis of LX2 cells.Realtime fluorescent quantitative PCR and Western blotting were used to analyze the effects of miR-34a-5p overexpression on sonic hedgehog(Shh),glioma-related oncogenes 1(Gli1),glioma-related oncogenes 2(Gli2),collagen typeⅠ(Col-Ⅰ),andα-smooth muscle actin(α-SMA)mRNA and protein expression.Results After LX2 cells were activated using TGF-β1,the cell viability was significantly increased and the expressions of Shh,Gli1,Gli2,Col-Ⅰ,andα-SMA was increased(all P<0.01).The specific inhibitor of the Hh signaling pathway cyclopamine reduced LX2 viability and down-regulated the expressions of Shh,Gli1,Gli2,Col-Ⅰ,andα-SMA proteins(all P<0.01).Overexpression of miR-34a-5p significantly increased the viability of the LX2 cells and the protein expression levels of Gli1,Gli2,Col-Ⅰ,andα-SMA(all P<0.05).Conclusion miR-34a-5p targeted the Hh signaling pathway in LX2 cells.Overexpression of miR-34a-5p up-regulates the expression of Hh signaling pathway-related proteins and promotes the activation of LX2 cells.