Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia.

Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia. Research Abstract Details 

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Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia. Abstract Text:

Ivana Cecic,Denise A Chan,Patrick D Sutphin,Pritha Ray,Sanjiv Sam Gambhir,Amato J Giaccia,Edward E Graves,Ivana Cecic,Denise A Chan,Patrick D Sutphin,Pritha Ray,Sanjiv Sam Gambhir,Amato J Giaccia,Edward E Graves,

Reporter gene techniques have been applied toward studying the physiologic phenomena associated with tumor hypoxia, a negative prognostic indicator. The purpose of this study was to assess the potential adverse effects of hypoxic conditions on the effectiveness of four commonly used reporter genes: Renilla luciferase, monomeric red fluorescent protein, thymidine kinase, and lacZ. Tumor-forming A375 cells expressing a trifusion reporter consisting of Renilla luciferase, monomeric red fluorescent protein, and thymidine kinase were subjected to decreasing oxygen tensions and assayed for reporter expression and activity. A375 cells expressing beta-galactosidase were similarly exposed to hypoxia, with activity of the reporter monitored by cleavage of the fluorescent substrate 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)-beta-galactoside (DDAOG). Generation of signal in in vivo tumor models expressing bioluminescent or beta-galactosidase reporters were also examined over the course of hypoxic stresses, either by tumor clamping or the antivascular agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA). Our findings indicate that bioluminescent and fluorescent reporter activity are decreased under hypoxia despite minimal variations in protein production, whereas beta-galactosidase reporter activity per unit protein was unchanged. These results demonstrate that combining beta-galactosidase with the DDAOG optical probe may be a robust reporter system for the in vivo study of tumor hypoxia.

Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia. Publishing Authors By Initials

I Cecic,DA Chan,PD Sutphin,P Ray,SS Gambhir,AJ Giaccia,EE Graves,I Cecic,DA Chan,PD Sutphin,P Ray,SS Gambhir,AJ Giaccia,EE Graves,

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Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Molecular imaging : official journal of the Societ

VOLUME: 6

Page Numbers: 219-28

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ISSN: 1535-3508

DAY: 22

MONTH: 08

YEAR: 2007

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LANGUAGE: eng

NlmUniqueID: 101120118

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Grant and Affiliation Information for Oxygen sensitivity of reporter genes: implications for preclinical imaging of tumor hypoxia.

AFFILIATION: Department of Radiation Oncology and Molecular Imaging Program, Stanford, Stanford University School of Medicine, Stanford, CA 94305-5847, USA.

Country: United States

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MEDLINETA: Mol Imaging

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