海南医学院学报2024，Vol.30Issue(24)：1859-1868,10.DOI:10.13210/j.cnki.jhmu.20240709.001
miR-21-5p靶向调控Skp2促进骨关节炎软骨细胞凋亡并抑制自噬
miR-21-5p targeted regulation of Skp2 promotes apoptosis and inhibits autophagy in osteoarthritis chondrocytes
摘要
Abstract
Objective:To explore the effect of miR-21-5p targeted regulation of Skp2 on apoptosis and autophagy in OA chon-drocytes from the perspective of human chondrocytes.Methods:TargetScan online website predicted the binding site of miR-21-5p and Skp2,the direct interaction between miR-21-5p and Skp2 was verified by dual luciferase assay,primary human chondro-cytes were selected for isolation and culture,cell proliferation ability was detected by CCK-8 assay,the effect of miR-21-5p on Skp2 in human chondrocytes,Skp2,Bax,Bcl-2,LC3Ⅰ,LC3Ⅱ,Beclin1,and CARM1 mRNA expression in human chondro-cytes was detected by qPCR,and Western blot was used to detect the protein expression level of Skp2,CARM1,Bax,Bcl-2,LC3Ⅱ/Ⅰ,Beclin1,and the construction of Skp2 overexpression vector at a later stage to further detect the expression of related proteins.Results:TargetScan online website predicted that there was a potential miR-21-5p binding sequence in the 3'UTR of Skp2 mRNA;dual-luciferase method verified that there was an interactions between miR-21-5p and Skp2;qPCR detected suc-cessful Skp2 overexpression validation,confirming that Skp2 was under-expressed in chondrocytes;Western blot assay showed that miR-21-5p could promote apoptosis and reduce autophagy in human chondrocytes by regulating the levels of Skp2,CARM1,Bax,Bcl-2,LC3Ⅱ/Ⅰ,and Beclin1.Conclusion:miR-21-5p negatively regulates Skp2,promotes apoptosis,and inhibits autoph-agy in OA chondrocytes.关键词
骨关节炎/软骨细胞/miR-21-5p/Skp2/凋亡/自噬Key words
Osteoarthritis/Chondrocytes/miR-21-5p/Skp2/Apoptosis/Autophagy分类
医药卫生引用本文复制引用
袁长深,李彦宏,刘晋邑,袁景钊,梅其杰,徐文飞,郭锦荣,段戡,莫坚..miR-21-5p靶向调控Skp2促进骨关节炎软骨细胞凋亡并抑制自噬[J].海南医学院学报,2024,30(24):1859-1868,10.基金项目
This study was supported by the National Natural Science Foundation of China(82060875,82160912),Natural Science Foundation of Guangxi Province(2023GXNSFAA026051),and Innovation Project of Guangxi Graduate Education of GXUCM(YCSW2024401) 国家自然科学基金资助项目(82060875,82160912) （82060875,82160912）
广西自然科学基金(2023GXNSFAA026051) （2023GXNSFAA026051）
广西中医药大学研究生教育创新计划项目(YCSW2024401) （YCSW2024401）
