Expression and function of Ets-1 during experimental acute renal failure in rats

Hiroyuki Tanaka, Yoshio Terada, Takahiko Kobayashi, Tomokazu Okado, Seiji Inoshita, Michio Kuwahara, Arun Seth, Yasufumi Sato, Sei Sasaki

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Abstract

The Ets family of transcription factors is defined by a conserved DNA-binding Ets domain that forms a winged helix-turn-helix structure motif. The Ets family is involved in a diverse array of biologic functions, including cellular growth, migration, and differentiation. The hypothesis in this study was that Ets-1 is re-expressed during regeneration after acute renal failure (ARF) and plays a key role in the transcriptional regulation of cyclin D1 and the cell cycle progression in renal tubular cells. For clarifying the significance of Ets-1 in ARF, a rat ARF model in vivo and LLC-PK1 cells as an in vitro model were used. After the left rat renal artery was clamped for 1 h, the whole kidney homogenate was examined and total RNA was extracted at 6, 12, 24, 48, and 72 h after reperfusion by Western blot analysis and real-time reverse transcription-PCR. Ets-1 mRNA and protein expression were strongly increased at 6 to 24 h after the ischemia, respectively. The expression of hypoxia-inducible factor-1α was increased dramatically as early as 6 h after ischemia-reperfusion and decreased at 48 and 72 h after ischemia-reperfusion. In the immunohistologic examination, Ets-1 was expressed in the proximal tubules and coexpressed with proliferating cell nuclear antigen (PCNA). Furthermore, overexpression of Ets-1 promoted the cell cycle and increased the promoter activity and protein expression of cyclin Dl in LLC-PK1 cells. Ets-1 promoter activity increased between 3 and 6 h in hypoxia, and hypoxia also induced changes in the Ets-1 protein level in LLC-PK1 cells. The Ets-1 induction by hypoxia was abolished by the transfection of dominant-negative hypoxia-inducible factor-1α. A gel shift assay demonstrated that Ets-1 binds to the ets-1 binding site of the cyclin D1 promoter in the ischemia-reperfusion condition. Overexpression of Ets-1 did not significantly change the caspase 3 activity or the value of cell death ELISA in LLC-PK1 cells. Taken together, these data suggest that Ets-1 plays a key role in the cell-cycle progression of renal tubules in ARF. The Ets-1 pathway may regulate the transcription of cyclin D1 and control the regeneration of renal tubules in ARF.

Original languageEnglish
Pages (from-to)3083-3092
Number of pages10
JournalJournal of the American Society of Nephrology
Volume15
Issue number12
DOIs
Publication statusPublished - 2004 Dec 1

ASJC Scopus subject areas

  • Nephrology

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    Tanaka, H., Terada, Y., Kobayashi, T., Okado, T., Inoshita, S., Kuwahara, M., Seth, A., Sato, Y., & Sasaki, S. (2004). Expression and function of Ets-1 during experimental acute renal failure in rats. Journal of the American Society of Nephrology, 15(12), 3083-3092. https://doi.org/10.1097/01.ASN.0000145459.54236.D3