miR-21-5p targeted regulation of Skp2 promotes apoptosis and inhibits autophagy in osteoarthritis chondrocytes
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.
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