海南医科大学学报2026，Vol.32Issue(9)：649-660,12.DOI:10.13210/j.cnki.jhmu.20250613.001
miR-296-3p通过靶向ZBTB20抑制HSC-T6细胞的活化
miR-296-3p inhibits the avtivation of HSC-T6 cell by targeting ZBTB20
摘要
Abstract
Objective:To investigate the role of miR-296-3p in the regulation of HSC-T6 activation in rat hepatic stellate cells by targeting zinc finger and BTB domain-containing protein 20(ZBTB20).Methods:The expression levels of fibrosis markers and miR-296-3p in rat hepatic stellate cells HSC-T6 stimulated by transforming growth factor-β1(TGF-β1)were detected by qRT-PCR.The miR-296-3p mimic/inhibitor and their respective controls were transfected into HSC-T6 cells using liposome transfec-tion,respectively.There were mimic NC group,miR-296-3p mimic group,inhibitor group and miR-296-3p inhibitor group.The effects of miR-296-3p on the activation ability of HSC-T6 cells were detected by qRT-PCR,Western Blot,CCK-8,colony forma-tion,and Transwell assays.After transfection with miR-296-3p mimic/inhibitor,the candidate target genes of miR-296-3p were predicted by bioinformatics methods and validated with dual luciferase gene reporter assay.qRT-PCR and Western Blot experi-ments were performed to detect the binding relationship between miR-296-3p and the candidate target gene ZBTB20.qRT-PCR,CCK-8 and Transwell assays were performed to analyse whether pcDNA3.1-ZBTB20 could reverse the inhibitory effect of miR-296-3p+mimics on the activation ability of HSC-T6 cells.Results:HSC-T6 cells were further activated by TGF-β1 stimulation,and the expression level of miR-296-3p was down-regulated in activated cells.miR-296-3p inhibited the proliferation and migration of HSC-T6,and decreased the expression level of hepatic fibrosis markers,collagen type I(Col1A1)and α-smooth muscle actin(α-SMA).The bioinformatics database prediction and the verification of the double luciferase gene reporter assay showed that ZBTB20 was a functional target gene of miR-296-3p and that miR-296-3p negatively regulates ZBTB20 expression in HSC-T6.ZBTB20 could partially reverse the inhibitory effect of miR-296-3p on HSC-T6 activation.Conclusion:miR-296-3p inhibits fur-ther activation of HSC-T6 cells by targeting and inhibiting ZBTB20 expression.关键词
miR-296-3p/ZBTB20/HSC-T6细胞/肝纤维化/增殖/迁移Key words
MiR-296-3p/ZBTB20/HSC-T6 cells/Hepatic fibrosis/Proliferation/Migration分类
医药卫生引用本文复制引用
周静,李以恒,李尹凡,崔笑妍,王梅梅,张荣花,刘志勇,章广玲..miR-296-3p通过靶向ZBTB20抑制HSC-T6细胞的活化[J].海南医科大学学报,2026,32(9):649-660,12.基金项目
This study was supported by the Hebei Province Natural Science Foundation Project(H2023209047) （H2023209047）
Hebei Provincial Department of Education Funded Project for Cultivating Innovative Ability of Graduate Students(CXZZSS2024056) 河北省自然科学基金资助项目(H2023209047) （CXZZSS2024056）
河北省教育厅在读研究生创新能力培养资助项目(CXZZSS2024056) （CXZZSS2024056）
