Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Green fluorescent protein is a suitable reporter of tumor hypoxia despite an oxygen requirement for chromophore formation
Autore:
Vordermark, D; Shibata, T; Brown, JM;
Indirizzi:
Stanford Univ, Sch Med, Dept Radiat Oncol, Div Radiat & Canc Biol, Stanford, CA 94305 USA Stanford Univ Stanford CA USA 94305 t & Canc Biol, Stanford, CA 94305 USA
Titolo Testata:
NEOPLASIA
fascicolo: 6, volume: 3, anno: 2001,
pagine: 527 - 534
SICI:
1522-8002(200111/12)3:6<527:GFPIAS>2.0.ZU;2-2
Fonte:
ISI
Lingua:
ENG
Soggetto:
PREDICTS RADIATION RESPONSE; CELLS; GENE; RADIOSENSITIZATION; PRETREATMENT; MISONIDAZOLE; CARCINOMA; CERVIX; CANCER; HEAD;
Keywords:
green fluorescent protein; hypoxia; transfection; reoxygenation; fibrosarcoma;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
29
Recensione:
Indirizzi per estratti:
Indirizzo: Brown, JM Stanford Univ, Sch Med, Dept Radiat Oncol, Div Radiat & Canc Biol, CCSR S,Room 1255,269 Campus Dr, Stanford, CA 94305 USA Stanford Univ CCSR S,Room 1255,269 Campus Dr Stanford CA USA 94305
Citazione:
D. Vordermark et al., "Green fluorescent protein is a suitable reporter of tumor hypoxia despite an oxygen requirement for chromophore formation", NEOPLASIA, 3(6), 2001, pp. 527-534

Abstract

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 hypoxiawas 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, and 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 <0.02% oxygen, but was unaffected by higher oxygen concentrations. Following 12 hours at <0.02% oxygen, cells transfected with the hypoxia-responsive vector exhibited maximum fluorescence after 4 hours of subsequent reoxygenation, reaching 68 +/-2% of thelevels in CMV promoter controls under aerobic conditions. With such reoxygenation, these cells exhibited a constant increase in fluorescence between 2% and <0.02% oxygen. EGFP chromophore formation is only affected by near-anoxic oxygen concentrations. The correlation of fluorescence and oxygen concentration is restored by a 4-hour reoxygenation period due to oxidation ofpre-synthesized EGFP and a delayed increase in EGFP protein synthesis.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 03/04/20 alle ore 10:58:34