Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor Hypoxia
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:...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publications
2007-07-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.2310/7290.2007.00017 |
| _version_ | 1797282071608033280 |
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| author | Ivana Cecic Denise A. Chan Patrick D. Sutphin Pritha Ray Sanjiv Sam Gambhir Amato J. Giaccia Edward E. Graves |
| author_facet | Ivana Cecic Denise A. Chan Patrick D. Sutphin Pritha Ray Sanjiv Sam Gambhir Amato J. Giaccia Edward E. Graves |
| author_sort | Ivana Cecic |
| collection | DOAJ |
| description | 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 β-galactosidase were similarly exposed to hypoxia, with activity of the reporter monitored by cleavage of the fluorescent substrate 7-hydroxy-9 H -(1, 3-dichloro-9, 9-dimethylacridin-2-one)-β-galactoside (DDAOG). Generation of signal in in vivo tumor models expressing bioluminescent or β-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 β-galactosidase reporter activity per unit protein was unchanged. These results demonstrate that combining β-galactosidase with the DDAOG optical probe may be a robust reporter system for the in vivo study of tumor hypoxia. |
| first_indexed | 2024-03-07T17:06:38Z |
| format | Article |
| id | doaj.art-37a614026c8741e687a0d4ee46a0fec2 |
| institution | Directory Open Access Journal |
| issn | 1536-0121 |
| language | English |
| last_indexed | 2024-03-07T17:06:38Z |
| publishDate | 2007-07-01 |
| publisher | SAGE Publications |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj.art-37a614026c8741e687a0d4ee46a0fec22024-03-03T02:32:19ZengSAGE PublicationsMolecular Imaging1536-01212007-07-01610.2310/7290.2007.0001710.2310_7290.2007.00017Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor HypoxiaIvana CecicDenise A. ChanPatrick D. SutphinPritha RaySanjiv Sam GambhirAmato J. GiacciaEdward E. GravesReporter 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 β-galactosidase were similarly exposed to hypoxia, with activity of the reporter monitored by cleavage of the fluorescent substrate 7-hydroxy-9 H -(1, 3-dichloro-9, 9-dimethylacridin-2-one)-β-galactoside (DDAOG). Generation of signal in in vivo tumor models expressing bioluminescent or β-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 β-galactosidase reporter activity per unit protein was unchanged. These results demonstrate that combining β-galactosidase with the DDAOG optical probe may be a robust reporter system for the in vivo study of tumor hypoxia.https://doi.org/10.2310/7290.2007.00017 |
| spellingShingle | Ivana Cecic Denise A. Chan Patrick D. Sutphin Pritha Ray Sanjiv Sam Gambhir Amato J. Giaccia Edward E. Graves Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor Hypoxia Molecular Imaging |
| title | Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor Hypoxia |
| title_full | Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor Hypoxia |
| title_fullStr | Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor Hypoxia |
| title_full_unstemmed | Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor Hypoxia |
| title_short | Oxygen Sensitivity of Reporter Genes: Implications for Preclinical Imaging of Tumor Hypoxia |
| title_sort | oxygen sensitivity of reporter genes implications for preclinical imaging of tumor hypoxia |
| url | https://doi.org/10.2310/7290.2007.00017 |
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