P53 upregulated by HIF-1α promotes hypoxia-induced G2/M arrest and renal fibrosis in vitro and in vivo

Limin Liu, Peng Zhang, Ming Bai, Lijie He, Lei Zhang, Ting Liu, Zhen Yang, Menglu Duan, Minna Liu, Baojian Liu, Rui Du, Qi Qian, Shiren Sun

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6 Scopus citations

Abstract

Hypoxia plays an important role in the genesis and progression of renal fibrosis. The underlying mechanisms, however, have not been sufficiently elucidated. We examined the role of p53 in hypoxia-induced renal fibrosis in cell culture (human and rat renal tubular epithelial cells) and a mouse unilateral ureteral obstruction (UUO) model. Cell cycle of tubular cells was determined by flow cytometry, and the expression of profibrogenic factors was determined by RT-PCR, immunohistochemistry, and western blotting. Chromatin immunoprecipitation and luciferase reporter experiments were performed to explore the effect of HIF-1α on p53 expression. We showed that, in hypoxic tubular cells, p53 upregulation suppressed the expression of CDK1 and cyclins B1 and D1, leading to cell cycle (G2/M) arrest (or delay) and higher expression of TGF-β, CTGF, collagens, and fibronectin. p53 suppression by siRNA or by a specific p53 inhibitor (PIF-α) triggered opposite effects preventing the G2/M arrest and profibrotic changes. In vivo experiments in the UUO model revealed similar antifibrotic results following intraperitoneal administration of PIF-α (2.2 mg/kg). Using gain-of-function, loss-of-function, and luciferase assays, we further identified an HRE3 region on the p53 promoter as the HIF-1α-binding site. The HIF-1α-HRE3 binding resulted in a sharp transcriptional activation of p53. Collectively, we show the presence of a hypoxia-activated, p53-responsive profibrogenic pathway in the kidney. During hypoxia, p53 upregulation induced by HIF-1α suppresses cell cycle progression, leading to the accumulation of G2/M cells, and activates profibrotic TGF-β and CTGF-mediated signaling pathways, causing extracellular matrix production and renal fibrosis.

Original languageEnglish (US)
Article numbermjy042
Pages (from-to)371-382
Number of pages12
JournalJournal of Molecular Cell Biology
Volume11
Issue number5
DOIs
StatePublished - Jan 1 2019

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Keywords

  • cell cycle (G2/M) arrest
  • HIF-1α
  • hypoxia
  • p53
  • renal tubulointerstitial fibrosis
  • TGF-β

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Liu, L., Zhang, P., Bai, M., He, L., Zhang, L., Liu, T., Yang, Z., Duan, M., Liu, M., Liu, B., Du, R., Qian, Q., & Sun, S. (2019). P53 upregulated by HIF-1α promotes hypoxia-induced G2/M arrest and renal fibrosis in vitro and in vivo. Journal of Molecular Cell Biology, 11(5), 371-382. [mjy042]. https://doi.org/10.1093/jmcb/mjy042