Source:http://linkedlifedata.com/resource/pubmed/id/15802864
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
2005-4-1
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pubmed:abstractText |
Hypoxia-inducible factor-1 (HIF-1) is responsible for various gene expressions related to tumor malignancy, such as metastasis, invasion and angiogenesis. Therefore, monitoring HIF-1 activity in solid tumors is becoming increasingly important in clinical and basic studies. To establish a convenient system for visualizing HIF-1 activity in tumor xenografts, we employed a promoter consisting of five copies of hypoxia response elements (5HRE), whose activity depends on HIF-1, and used a derivative of green fluorescence protein (d2EGFP) as a reporter gene. A human melanoma cell line, Be11, which contains the 5HRE-d2EGFP gene, showed fluorescence in response to hypoxia. The fluorescent intensity correlated inversely with the surrounding oxygen tension, and was time-dependent for the hypoxic treatment. Reoxygenation resulted in a rapid decrease in fluorescence due to the signal sequence for protein degradation encoded in d2EGFP, which enabled monitoring of HIF-1 activity in real-time. Heterogeneous fluorescence was observed in the solid tumor of a non-sacrificed tumor-bearing mouse. Immunohistochemical analysis confirmed that d2EGFP-expressing regions overlapped with the ones stained with a hypoxia marker, pimonidazole. These results suggest that the 5HRE-d2EGFP gene is suitable for the real-time imaging of HIF-1-activating cells in vivo, due to the short half-life of the d2EGFP protein as well as the specificity of the 5HRE promoter for HIF-1 activity.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/HIF1A protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Hypoxia-Inducible Factor 1,
http://linkedlifedata.com/resource/pubmed/chemical/Hypoxia-Inducible Factor 1, alpha...,
http://linkedlifedata.com/resource/pubmed/chemical/Nuclear Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Transcription Factors,
http://linkedlifedata.com/resource/pubmed/chemical/Tumor Markers, Biological
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0449-3060
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
46
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
93-102
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:15802864-Cell Hypoxia,
pubmed-meshheading:15802864-Cell Line, Tumor,
pubmed-meshheading:15802864-Computer Systems,
pubmed-meshheading:15802864-DNA-Binding Proteins,
pubmed-meshheading:15802864-Flow Cytometry,
pubmed-meshheading:15802864-Humans,
pubmed-meshheading:15802864-Hypoxia-Inducible Factor 1,
pubmed-meshheading:15802864-Hypoxia-Inducible Factor 1, alpha Subunit,
pubmed-meshheading:15802864-Melanoma,
pubmed-meshheading:15802864-Microscopy, Fluorescence,
pubmed-meshheading:15802864-Nuclear Proteins,
pubmed-meshheading:15802864-Transcription Factors,
pubmed-meshheading:15802864-Transplantation, Heterologous,
pubmed-meshheading:15802864-Tumor Markers, Biological
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pubmed:year |
2005
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pubmed:articleTitle |
Real-time imaging of hypoxia-inducible factor-1 activity in tumor xenografts.
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pubmed:affiliation |
Department of Therapeutic Radiology and Oncology, Kyoto University Graduate School of Medicine, Kyoto, Japan.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Evaluation Studies
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