Rodent-specific hypoxia response elements enhance PAI-1 expression through HIF-1 or HIF-2 in mouse hepatoma cells

  • Authors:
    • Yong-Tae Ahn
    • Mei-Sze Chua
    • James P. Whitlock
    • Yong-Chil Shin
    • Woon-Heung Song
    • Yongkuk Kim
    • Chi-Yong Eom
    • Won Gun An
  • View Affiliations

  • Published online on: December 1, 2010     https://doi.org/10.3892/ijo_00000817
  • Pages: 1627-1638
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Abstract

Plasminogen activator inhibitor-1 (PAI-1) is an important regulator of numerous pathophysiological processes such as inflammation, thrombosis, angiogenesis and tumor metastasis. Its expression is induced by hypoxia at the transcriptional level, via the hypoxia inducible factor-1 (HIF-1) or -2 (HIF-2). In this study, we elucidated the mechanism of transcriptional regulation of mouse PAI-1 gene by hypoxia in mouse hepatoma cells. We searched for hypoxia response elements (HREs) of murine PAI-1 promoter using several molecular biological assays. DNAse I hypersensitivity assay first suggested that PAI-1 gene expression is up-regulated by protein-DNA interactions at the -3.6- and -3-kb upstream regions of the PAI-1 gene transcription start site. An approximately 6.4-kb region of DNA containing the 5'-flanking promoter region of the PAI-1 gene was isolated, mapped, and cloned into reporter gene assay vectors and sequenced. Luciferase reporter gene assay subsequently identified two functional HREs, located around -3.6 kb of the 5'-flanking promoter region of PAI-1 gene that were responsible for the enhancement of luciferase reporter gene activity. Mutation of the HREs in this fragment abolished luciferase reporter gene activity. Finally, in vitro and in vivo protein-DNA interaction assays confirmed binding of the two HREs to HIF-1 or HIF-2 protein. Our results show that two HREs located around -3.6 kb of the 5'-flanking promoter region of the mouse PAI-1 gene function as hypoxia enhancers, which, alongside other regulatory regions, control PAI-1 gene transcription by HIF-1 or HIF-2 under hypoxic environments in mouse hepatoma cells.

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December 2010
Volume 37 Issue 6

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Ahn Y, Chua M, Whitlock JP, Shin Y, Song W, Kim Y, Eom C and An WG: Rodent-specific hypoxia response elements enhance PAI-1 expression through HIF-1 or HIF-2 in mouse hepatoma cells. Int J Oncol 37: 1627-1638, 2010
APA
Ahn, Y., Chua, M., Whitlock, J.P., Shin, Y., Song, W., Kim, Y. ... An, W.G. (2010). Rodent-specific hypoxia response elements enhance PAI-1 expression through HIF-1 or HIF-2 in mouse hepatoma cells. International Journal of Oncology, 37, 1627-1638. https://doi.org/10.3892/ijo_00000817
MLA
Ahn, Y., Chua, M., Whitlock, J. P., Shin, Y., Song, W., Kim, Y., Eom, C., An, W. G."Rodent-specific hypoxia response elements enhance PAI-1 expression through HIF-1 or HIF-2 in mouse hepatoma cells". International Journal of Oncology 37.6 (2010): 1627-1638.
Chicago
Ahn, Y., Chua, M., Whitlock, J. P., Shin, Y., Song, W., Kim, Y., Eom, C., An, W. G."Rodent-specific hypoxia response elements enhance PAI-1 expression through HIF-1 or HIF-2 in mouse hepatoma cells". International Journal of Oncology 37, no. 6 (2010): 1627-1638. https://doi.org/10.3892/ijo_00000817