Author/Editor     Coralli, Claudia; Čemažar, Maja; Kanthou, Chryso; Tozer, Gillian M; Dachs, Gabi U
Title     Limitations of the reporter green fluorescent protein under simulated tumor conditions
Type     članek
Source     Cancer Res
Vol. and No.     Letnik 61, št. 12
Publication year     2001
Volume     str. 4784-90
Language     eng
Abstract     This paper reports a detailed analysis of the effect of low oxygen conditions (hypoxia) on the reporter green tiuorescent protein (GFP). It questions the feasibility of using GFP for gene expression studies under tumor conditions. Hypoxia is a characteristic of both experimental and clinical tumors. Several important factors are pointed out which need to be considered when using GFP as reporter gene. GFP tluorescence is the final product of a long and complex pathway involving transcription, translation, and posttranslational modifications. All of these steps may be affected by the availability of oxygen. We show specifically that cellular GFP fluorescence decreased with reduced oxygenation, anoxia virtually eliminated tluorescence and protein levels, and fluorescence recovery after anoxia required 5-10 h of reoxygenation. In conclusion, GFP appears to be a good marker gene to study location or movement of proteins or cells but should be used with great caution as a reporter of gene expression under tumor conditions.
Descriptors     BLADDER NEOPLASMS
GENES, REPORTER
LUMINESCENT PROTEINS
OXYGEN
MICE, SCID
CELL HYPOXIA
FLUORESCENCE
GENE EXPRESSION
INDICATORS AND REAGENTS
RNA, MESSENGER
TRANSFECTION
TUMOR CELLS, CULTURED