四川医学2024，Vol.45Issue(7)：709-715,7.DOI:10.16252/j.cnki.issn1004-0501-2024.07.003
miR-30a-5p靶向JAK1调控人胃腺癌AGS细胞自噬的机制研究
Mechanism of miR-30a-5p Targeting JAK1 to Regulate Autophagy in Human Gastric Adenocarcinoma AGS Cells
摘要
Abstract
Objective To explore the mechanism by which miR-30a-5p targets JAK1 to regulate autophagy in human gas-tric adenocarcinoma AGS cells.Methods Human normal gastric mucosal epithelial cells(GES-1)and gastric adenocarcinoma cells(AGS)were cultured in vitro.RT-qPCR was performed to detect the expression level of miR-30a-5p.The Starbase database was used to predict the expression level of miR-30a-5p and its potential binding site to JAK1,and dual-luciferase reporter assay was performed to verify the target-regulatory effect of miR-30a-5p on JAK1.Immunofluorescence was conducted to detect the level of LC3-positive cells for autophagy markers,and WB was utilized to detect the expression level of JAK1 and autophagy-related pro-teins(Beclin-1,LC3 Ⅱ/Ⅰ,and p62).Results The expression level of miR-30a-5p in the AGS group was significantly lower than that in the GES-1 group,with significantly lower expression levels of autophagy-related proteins(Beclin-1 and LC3 Ⅱ/Ⅰ)and the autophagy marker LC3 and markedly higher expression levels of p62 protein.miR-30a-5p had a binding site to JAK1 and targeted JAK1 negatively.Finally,rescue experiments displayed that compared with the miR-30a-5p mimics+oe-NC group,the miR-30a-5p mimics+oe-JAKl group exhibited substantial increases in the expression level of autophagy-related proteins(Bec-lin-1 and LC3 Ⅱ/Ⅰ)and significant reductions in p62 protein expression in AGS cells.Further immunofluorescence also revealed a significant increase in LC3 fluorescence intensity after miR-30a-5p overexpression plus JAK1 overexpression in AGS cells.Con-clusion miR-30a-5p targets and negatively regulates JAK1 protein expression and inhibits autophagy in human gastric adenocarci-noma AGS cells.关键词
胃腺癌/miR-30a-5p/JAK1/AGS细胞/细胞自噬Key words
gastric adenocarcinoma/miR-30a-5p/JAK1/AGS cells/autophagy分类
医药卫生引用本文复制引用
刘海英,曹天柱,崔自重..miR-30a-5p靶向JAK1调控人胃腺癌AGS细胞自噬的机制研究[J].四川医学,2024,45(7):709-715,7.基金项目
长沙市自然科学基金(编号:kq2202014) （编号:kq2202014）
