Knockdown of circRNA WD repeat containing protein 1 inhibits proliferation and induces apoptosis of chondrocytes in knee osteoarthritis

doi:10.12307/2024.323

Abstract

Abstract: BACKGROUND: Circular RNAs (circRNAs) play important roles in a variety of diseases or tumors, and recent findings have revealed that circRNAs are abnormally expressed in knee osteoarthritis and are involved in disease progression through microRNA/mRNA regulation.
OBJECTIVE: To investigate the effect of circRNA WD repeat containing protein 1 (circ-BRWD1)/miR-488-3p/DNA methyltransferase 3A (DNMT3A) on the proliferation and apoptosis of chondrocytes in knee osteoarthritis.
METHODS: Real-time fluorescence quantitative PCR was performed to detect the expression of circ-BRWD1, miR-488-3p, DNMT3A in knee osteoarthritis chondrocytes. Cells were divided into si-NC group, si-circ-BRWD1 group, vector group, circ-BRWD1 group, si-circ-BRWD1+anti-miR-NC group, si-circ-BRWD1+anti-miR-488-3p group, miR-NC group, miR-488-3p group, anti-miR-NC group, anti-miR-488-3p group, miR-488-3p+vector group, miR 488-3p+DNMT3A group. Real-time fluorescence quantitative PCR was used to detect circ-BRWD1, miR-488-3p, DNMT3A expression, MTT and flow cytometry assay were used to detect cell proliferation and apoptosis. Western blot assay was used to detect DNMT3A and proliferation/apoptosis-related protein expression. Dual luciferase reporter assay was used to Dual luciferase reporter assay to detect the targeting relationship of circ-BRWD1 with miR-488-3p and miR-488-3p with DNMT3A.
RESULTS AND CONCLUSION: circ-BRWD1 and DNMT3A were highly expressed and miR-488-3p was lowly expressed in knee osteoarthritis chondrocytes compared with normal chondrocytes. Knockdown of circ-BRWD1 or overexpression of miR-488-3p inhibited proliferation and induced apoptosis in knee osteoarthritis chondrocytes. circ-BRWD1 targeted negative regulation of miR-488-3p and inhibition of miR-488-3p reversed the effect of circ-BRWD1 knockdown on chondrocyte proliferation and apoptosis in knee osteoarthritis. miR-488-3p targeted negative regulation of DNMT3A and upregulation of DNMT3A reversed the effect of miR-488-3p overexpression on chondrocyte proliferation and apoptosis in knee osteoarthritis. circ-BRWD1 could regulate the expression of DNMT3A by regulating miR-488-3p. To conclude, knockdown of circ-BRWD1 inhibits chondrocyte proliferation and induces apoptosis in knee osteoarthritis, and the mechanism of action may be related to the regulation of miR-488-3p/DNMT3A axis.

Key words: circ-BRWD1, miR-488-3p, DNMT3A, knee osteoarthritis, chondrocyte, proliferation, apoptosis

CLC Number:

Shen Feiyan, Yao Jixiang, Su Shanshan, Zhao Zhongmin, Tang Weidong. Knockdown of circRNA WD repeat containing protein 1 inhibits proliferation and induces apoptosis of chondrocytes in knee osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(4): 499-504.

Figures/Tables 6

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