WWP2 ameliorates acute kidney injury by mediating p53 ubiquitylation and degradation

Abstract

E3 ubiquitin ligase gene, WWP2, is associated with acute kidney injury (AKI). This research was conducted to explore the role of WWP2 in AKI. AKI cell model was produced in human renal proximal tubular epithelial cell line (HK‐2) by ischemia‐reperfusion (IR) injury. CCK8 and flow cytometry assay were performed to explore the influence of WWP2 overexpression on cell proliferation and apoptosis of IR‐induced HK‐2 cells. Quantitative real‐time PCR and immunoblotting (IB) were performed to assess the gene and protein expression. Then, the influence of WWP2 on p53 ubiquitylation and degradation was estimated by immunoprecipitation assay. Our data indicated that WWP2 was down‐regulated and p53 was up‐regulated in IR‐induced HK‐2 cells. WWP2 overexpression promoted proliferation and inhibited apoptosis of IR‐induced HK‐2 cells. And WWP2 interacted with p53 and regulated p53 ubiquitylation and degradation. Furthermore, the influence of WWP2 on cell proliferation and apoptosis was rescued by MG132 (proteasome inhibitor) treatment. In conclusion, our work described for the first time the role of WWP2 in AKI, showing that WWP2 ameliorated AKI by mediating p53 ubiquitylation and degradation. Moreover, the study offers some important insights into the occurrence of AKI and WWP2 may be a novel target of AKI treatment.Significance of the studyOur data elaborates that WWP2 has protective effect against AKI by mediating p53 ubiquitylation and degradation. Thus, WWP2 might be a therapeutic target for AKI.

miR-328-5p Induces Human Intervertebral Disc Degeneration by Targeting WWP2

“…Xu et al found that WWP2 siRNA inhibited Bcl-2 expression by promoting Bax and Caspase7/8 to induce apoptosis of liver cancer cells [ 53 ]. Even more, overexpression of WWP2 could inhibit the apoptosis of human renal tubular epithelial cells by inducing Bcl-2 expressionand and inhibiting Bax expression [ 40 ]. However, the regulatory mechanism of miR-328-5p and its target gene WWP2 in IDD has not been reported.…”

“…The experiment was performed three times. Experiment is based on U6 and β -actin [ 40 ]. Gene expression was measured using the 2 −△△Ct .…”

“…After primary antibody incubation, the NC film was washed three times with TBST and addted to goat anti-rabbit or mouse antibody (Vicmed, Xuzhou, China) for 2 h at room temperature in the dark. Beta actin was selected as the internal control [40]. , c, and d) the correlation between miR-328-5p and its target genes.…”

NAT10‐mediated ac4C modification promotes ectoderm differentiation of human embryonic stem cells via acetylating NR2F1 mRNA

“…The protein stability assay was according to the previous reports 20–22 . Briefly, the DMSO‐treated NEP cells (Day 7) and RE‐treated NEP cells (Day 7) were treated with an inhibitor of protein synthesis, cycloheximide (CHX, 100 μg/mL; Sigma, USA).…”

“…The protein stability assay was according to the previous reports. 20 , 21 , 22 Briefly, the DMSO‐treated NEP cells (Day 7) and RE‐treated NEP cells (Day 7) were treated with an inhibitor of protein synthesis, cycloheximide (CHX, 100 μg/mL; Sigma, USA). The cells were obtained at indicated times for analysing the protein stability of NR2F1 by Western blotting.…”