Green Fluorescent Protein is a Suitable Reporter of Tumor Hypoxia Despite an Oxygen Requirement for Chromophore Formation
| Autor: | Dirk Vordermark, Toru Shibata, J. Martin Brown |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2001 |
| Předmět: | |
| Zdroj: | Neoplasia: An International Journal for Oncology Research, Vol 3, Iss 6, Pp 527-534 (2001) |
| Druh dokumentu: | article |
| ISSN: | 1476-5586 1522-8002 |
| DOI: | 10.1038/sj.neo.7900192 |
| Popis: | The oxygen requirement for chromophore formation potentially limits the use of green fluorescent protein as a reporter under hypoxic conditions. In the light of this, the applicability of a hypoxia-responsive enhanced green fluorescent protein (EGFP)-based system to the measurement of tumor hypoxia was tested in human HT 1080 fibrosarcoma cells stably transfected with a destabilized EGFP vector containing the hypoxia-responsive 5HRE-hCMVmp promoter or, as a positive control, the strong constitutive CMV promoter. After various schedules of hypoxia and reoxygenation, EGFP fluorescence of live cells was assessed by flow cytometry, protein levels were analyzed by Western blot. Fluorescence of CMV promoter positive control cells dropped to 38±5% of aerobic levels after 12 hours at |
| Databáze: | Directory of Open Access Journals |
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