Circ-FBXW12 aggravates the development of diabetic nephropathy by binding to miR-31-5p to induce LIN28B

• Published in: Diabetology and metabolic syndrome 2021-12, Vol.13 (1), p.1-141, Article 141
• Main Authors: Sun, Aidong, Sun, Ningshuang, Liang, Xiao, Hou, Zhenbo
• Format: Article
• Language: English
• Subjects: Analysis; Apoptosis; Binding sites; Blood diseases; Cell cycle; Cell growth; Cell proliferation; Cell viability; Cholecystokinin; Circ-FBXW12; Cytokines; Diabetes; Diabetes mellitus; Diabetic nephropathies; Diabetic nephropathy; Diabetic neuropathy; Enzyme-linked immunosorbent assay; Ewings sarcoma; Extracellular matrix; Flow cytometry; Glucose; HMCs; Immunoprecipitation; Inflammation; Kinases; LIN28B; Malondialdehyde; Mesangial cells; MicroRNA; MicroRNAs; miR-31-5p; miRNA; Nephropathy; Oxidative stress; Pathogenesis; Polymerase chain reaction; Proteins; Reverse transcription; Ribonuclease; Superoxide; Superoxide dismutase; Tumor necrosis factor-TNF; Type 2 diabetes
• ISSN: 1758-5996; 1758-5996
• DOI: 10.1186/s13098-021-00757-x

Background The involvement of circular RNAs (circRNAs) in diabetic nephropathy (DN) has been gradually identified. In this study, we aimed to explore the functions of circRNA F-box/WD repeat-containing protein 12 (circ-FBXW12) in DN development. Methods Reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay was performed for the levels of circ-FBXW12, FBXW12 mRNA, microRNA-31-5p (miR-31-5p) and Lin-28 homolog B (LIN28B) mRNA. RNase R assay was used to analyze the stability of circ-FBXW12. Cell Counting Kit-8 (CCK-8) assay, flow cytometry analysis and 5-ethynyl-2'- deoxyuridine (EdU) assay were employed to evaluate cell viability, cell cycle and proliferation, respectively. Enzyme linked immunosorbent assay (ELISA) was done to measure the concentrations of inflammatory cytokines. Western blot assay was conducted for protein levels. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were examined with commercial kits. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to verify the relationships among circ-FBXW12, miR-31-5p and LIN28B. Results Circ-FBXW12 level was increased in DN patients' serums and high glucose (HG)-induced human mesangial cells (HMCs). Circ-FBXW12 knockdown suppressed cell proliferation, arrested cell cycle, reduced extracellular matrix (ECM) production and oxidative stress in HG-induced HMCs. Circ-FBXW12 was identified as the sponge for miR-31-5p, which then directly targeted LIN28B. MiR-31-5p inhibition reversed circ-FBXW12 knockdown-mediated effects on cell proliferation, cell cycle process, ECM production and oxidative in HG-triggered HMCs. Moreover, miR-31-5p overexpression showed similar results with circ-FBXW12 knockdown in HG-stimulated HMC progression, while LIN28B elevation reversed the effects. Conclusion Circ-FBXW12 knockdown suppressed HG-induced HMC growth, inflammation, ECM accumulation and oxidative stress by regulating miR-31-5p/LIN28B axis. Keywords: DN, HMCs, Circ-FBXW12, miR-31-5p, LIN28B