Molecular Mechanism Study of Circular Ribonucleic Acid 0136474 in Regulating Proliferation and Apoptosis of Osteoarthritis Chondrocytes
Objective:To investigate the effect of circular RNA-0136474(circ_0136474)on the proliferation and apoptosis of osteoarthritis chondrocytes by regulating the micro RNA-127-5p(miR-127-5p)/Zest homolog 2(EZH2)axis and its molecular mechanism.Methods:The osteoarthritis patients were collected.The expression of circ_0136474 in cartilage tis-sue of osteoarthritis patients was detected;human chondrocytes CHON-001 were induced by IL-1β and divided into control group,IL-1β group,si-NC group,si-circ_0136474 group,si-circ_0136474+anti-miR-NC group,si-circ_0136474+anti-miR-127-5p group,miR-NC group,and miR-127-5p mimics group.The CCK-8 was applied to detect the cell viability,flow cytometer was applied to detect the cell cycle and apoptosis,RT-qPCR was applied to detect the expression of circ_0136474,miR-127-5p,EZH2,and proliferating cell nuclear antigen(PCNA)mRNA,ELISA kit was applied to detect the levels of IL-6,TNF-α,and MMP-13,Western Blot was applied to detect the expression of PCNA,Bcl-2,Bax,and EZH2 proteins,double luciferase reporter gene experiment was applied to verify the targeting relationship between miR-127-5p and circ_0136474,EZH2.Results:The circ_0136474 was highly expressed in cartilage tissue of osteoarthritis patients;after induction by IL-1β,the cell viability,S phase,PCNA ex-pression,Bcl-2 expression,and miR-127-5p were obviously reduced,the proportion of G0-G1 phase cells,cell apoptosis rate,Bax,IL-6,TNF-α,MMP-13,circ_0136474,EZH2 protein,and EZH2 mRNA were obviously increased(P＜0.05).The double luciferase reporter gene experiment showed that circ_0136474 and EZH2 had a targeting relationship with miR-127-5p,silencing circ_0136474 can promote cell proliferation,prolong the S phase,shorten the G0-G1 phase,inhibit cell apoptosis,reduce the levels of IL-6,TNF-α,and MMP-13(P＜0.05),inhibiting the expression of miR-127-5p can reverse the inhibitory effects of silencing circ_0136474 on cell apoptosis,IL-6,TNF-α,and MMP-13(P＜0.05).Conclu-sion:The circ_0136474 is highly expressed in osteoarthritis cartilage tissue,and silencing circ_0136474 can promote chon-drocyte proliferation and inhibit cell apoptosis by regulating the miR-127-5p/EZH2 axis.
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