AHR, a novel acute hypoxia-response sequence, drives reporter gene expression under hypoxia in vitro and in vivo

Mehmet Zeynel Cilek, Satoshi Hirohata, Omer Faruk Hatipoglu, Hiroko Ogawa, Toru Miyoshi, Junko Inagaki, Takashi Ohtsuki, Hiroshi Harada, Shigeshi Kamikawa, Shozo Kusachi, Yoshifumi Ninomiya

Research output: Contribution to journalArticle

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Abstract

ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1) is an early immediate gene. We have previously reported that ADAMTS1 was strongly induced by hypoxia. In this study, we investigated whether ADAMTS1 promoter-driven reporter signal is detectable by acute hypoxia. We constructed the GFP (green fluorescent protein) expression vector [AHR (acute hypoxia-response sequence)-GFP] under the control of ADAMTS1 promoter and compared it with the constitutive GFP-expressing vector under the control of CMV (cytomegalovirus promoter-GFP). We transduced AHR-GFP and examined whether GFP signals can be detected under the acute hypoxia. When the human umbilical vein [HUVEC (human umbilical vein endothelial cells)] was transduced under normoxia, there were few GFP signals, while CMVGFP showed considerable GFP signals. When HUVEC was stimulated with hypoxia, GFP signals from AHR-GFP gene were induced under hypoxic conditions. Notably, the GFP signals peaked at 3 h under hypoxia. In ischaemic hind limb model, transduced AHR-GFP showed hypoxic induction of GFP signals. In summary, we have demonstrated that the AHR system induced the reporter gene expression by acute hypoxia, and its induction is transient. This is the first report showing the unique acute hypoxia-activated gene expression system.

Original languageEnglish
Pages (from-to)1-8
Number of pages8
JournalCell Biology International
Volume35
Issue number1
DOIs
Publication statusPublished - Jan 2011

Fingerprint

Green Fluorescent Proteins
Reporter Genes
Gene Expression
Thrombospondin 1
Disintegrins
Metalloproteases
In Vitro Techniques
Hypoxia
Umbilical Veins
Immediate-Early Genes
Human Umbilical Vein Endothelial Cells
Cytomegalovirus
Extremities

Keywords

  • A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)
  • Hypoxia
  • Metalloproteinase
  • Promoter

ASJC Scopus subject areas

  • Cell Biology

Cite this

AHR, a novel acute hypoxia-response sequence, drives reporter gene expression under hypoxia in vitro and in vivo. / Cilek, Mehmet Zeynel; Hirohata, Satoshi; Hatipoglu, Omer Faruk; Ogawa, Hiroko; Miyoshi, Toru; Inagaki, Junko; Ohtsuki, Takashi; Harada, Hiroshi; Kamikawa, Shigeshi; Kusachi, Shozo; Ninomiya, Yoshifumi.

In: Cell Biology International, Vol. 35, No. 1, 01.2011, p. 1-8.

Research output: Contribution to journalArticle

Cilek, Mehmet Zeynel ; Hirohata, Satoshi ; Hatipoglu, Omer Faruk ; Ogawa, Hiroko ; Miyoshi, Toru ; Inagaki, Junko ; Ohtsuki, Takashi ; Harada, Hiroshi ; Kamikawa, Shigeshi ; Kusachi, Shozo ; Ninomiya, Yoshifumi. / AHR, a novel acute hypoxia-response sequence, drives reporter gene expression under hypoxia in vitro and in vivo. In: Cell Biology International. 2011 ; Vol. 35, No. 1. pp. 1-8.
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