The combination of hypoxia-response enhancers and an oxygen-dependent proteolytic motif enables real-time imaging of absolute HIF-1 activity in tumor xenografts

The transcriptional activity of hypoxia-inducible factor-1 (HIF-1) is associated with tumor malignancies; therefore, it is important to comprehend its dynamism in solid tumors. However, a molecular imaging strategy to accurately access it remains to be developed. We constructed here a novel HIF-1-de...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2007-09, Vol.360 (4), p.791-796
Main Authors: Harada, Hiroshi, Kizaka-Kondoh, Shinae, Itasaka, Satoshi, Shibuya, Keiko, Morinibu, Akiyo, Shinomiya, Kazumi, Hiraoka, Masahiro
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
ANOXIA
Base Sequence
Cell Hypoxia - drug effects
DNA Primers
GENES
Genes, Reporter
HeLa Cells
Humans
Hydrolysis
Hypoxia
Hypoxia-Inducible Factor 1, alpha Subunit - metabolism
Hypoxia-inducible factor-1 (HIF-1)
Hypoxia-response element (HRE)
LUCIFERASE
LUMINESCENCE
NEOPLASMS
Neoplasms - metabolism
Optical imaging
OXYGEN
Oxygen - metabolism
Oxygen-dependent degradation domain (ODD domain)
PROMOTERS
Transplantation, Heterologous
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Summary:The transcriptional activity of hypoxia-inducible factor-1 (HIF-1) is associated with tumor malignancies; therefore, it is important to comprehend its dynamism in solid tumors. However, a molecular imaging strategy to accurately access it remains to be developed. We constructed here a novel HIF-1-dependent reporter gene, 5HREp-ODD-luc, in which 5 copies of the hypoxia-response element (5HRE) enhance expression of the oxygen-dependent degradation (ODD) domain and luciferase (luc) fusion under hypoxia. Because the ODD domain caused the oxygen-dependent degradation of the ODD-Luc protein, the novel reporter gene showed little leak of luminescence under normoxia. Such a property caused an increase of the hypoxia-responsiveness up to about 4.7 × 10 4-fold. Moreover, the ODD domain caused rapid degradation of the ODD-Luc protein under normoxia, the luminescence reflected the dynamism of HIF-1 activity in real-time. The superiority of the novel reporter gene will surely accelerate analysis of the intratumoral HIF-1 activity during tumor progression and cancer treatments.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2007.06.149