Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of Action
The signaling pathway(s) and molecular target(s) for 5,6-dimethylxanthenone-4-acetic acid (DMXAA), a tumor vascular disrupting agent in late stages of clinical development, are still undefined. As an approach toward identifying potential targets for DMXAA, a tritiated azido-analog of DMXAA was used...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2010-09-01
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| Series: | Neoplasia: An International Journal for Oncology Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1476558610800776 |
| _version_ | 1818359039454609408 |
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| author | Romy Brauer Liang-Chuan S. Wang See-Tarn Woon David J.A. Bridewell Kimiora Henare Dieter Malinger Brian D. Palmer Stefanie N. Vogel Claudine Kieda Sofian M. Tijono Lai-Ming Ching |
| author_facet | Romy Brauer Liang-Chuan S. Wang See-Tarn Woon David J.A. Bridewell Kimiora Henare Dieter Malinger Brian D. Palmer Stefanie N. Vogel Claudine Kieda Sofian M. Tijono Lai-Ming Ching |
| author_sort | Romy Brauer |
| collection | DOAJ |
| description | The signaling pathway(s) and molecular target(s) for 5,6-dimethylxanthenone-4-acetic acid (DMXAA), a tumor vascular disrupting agent in late stages of clinical development, are still undefined. As an approach toward identifying potential targets for DMXAA, a tritiated azido-analog of DMXAA was used to probe for cellular binding proteins. More than 20 cytosolic proteins from murine splenocytes, RAW 264.7 cells, and the HECPP immortalized endothelial cells were photoaffinity-labeled. Although no protein domain, fold, or binding site for a specific ligand was found to be shared by all the candidate proteins, essentially all were noted to be oxidizable proteins, implicating a role for redox signaling in the action of DMXAA. Consistent with this hypothesis, DMXAA caused an increase in concentrations of reactive oxygen species (ROS) in RAW264.7 cells during the first 2 hours. This increase in ROS was suppressed in the presence of the antioxidant, N-acetyl-L-cysteine, which also suppressed DMXAA-induced cytokine production in the RAW 264.7 cells with no effects on cell viability. Short interfering RNA (siRNA)-mediated knockdown of one of the photoaffinity-labeled proteins, superoxide dismutase 1, an ROS scavenger, resulted in an increase in tumor necrosis factor-α production by RAW 264.7 cells in response to DMXAA compared with negative or positive controls transfected with nontargeting or lamin A/C-targeting siRNA molecules, respectively. The results from these lines of study all suggest that redox signaling plays a central role in cytokine induction by DMXAA. |
| first_indexed | 2024-12-13T20:38:33Z |
| format | Article |
| id | doaj.art-baf727a07c2246b887a6ea00cdd7b610 |
| institution | Directory Open Access Journal |
| issn | 1476-5586 1522-8002 |
| language | English |
| last_indexed | 2024-12-13T20:38:33Z |
| publishDate | 2010-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neoplasia: An International Journal for Oncology Research |
| spelling | doaj.art-baf727a07c2246b887a6ea00cdd7b6102022-12-21T23:32:13ZengElsevierNeoplasia: An International Journal for Oncology Research1476-55861522-80022010-09-0112975576510.1593/neo.10636Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of ActionRomy Brauer0Liang-Chuan S. Wang1See-Tarn Woon2David J.A. Bridewell3Kimiora Henare4Dieter Malinger5Brian D. Palmer6Stefanie N. Vogel7Claudine Kieda8Sofian M. Tijono9Lai-Ming Ching10Auckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandUniversity of Maryland, Baltimore, Baltimore, MD, USACentre de Biophysique Moleculaire, CNRS UPR4301, Orleans, FranceAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandAuckland Cancer Society Research Centre, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New ZealandThe signaling pathway(s) and molecular target(s) for 5,6-dimethylxanthenone-4-acetic acid (DMXAA), a tumor vascular disrupting agent in late stages of clinical development, are still undefined. As an approach toward identifying potential targets for DMXAA, a tritiated azido-analog of DMXAA was used to probe for cellular binding proteins. More than 20 cytosolic proteins from murine splenocytes, RAW 264.7 cells, and the HECPP immortalized endothelial cells were photoaffinity-labeled. Although no protein domain, fold, or binding site for a specific ligand was found to be shared by all the candidate proteins, essentially all were noted to be oxidizable proteins, implicating a role for redox signaling in the action of DMXAA. Consistent with this hypothesis, DMXAA caused an increase in concentrations of reactive oxygen species (ROS) in RAW264.7 cells during the first 2 hours. This increase in ROS was suppressed in the presence of the antioxidant, N-acetyl-L-cysteine, which also suppressed DMXAA-induced cytokine production in the RAW 264.7 cells with no effects on cell viability. Short interfering RNA (siRNA)-mediated knockdown of one of the photoaffinity-labeled proteins, superoxide dismutase 1, an ROS scavenger, resulted in an increase in tumor necrosis factor-α production by RAW 264.7 cells in response to DMXAA compared with negative or positive controls transfected with nontargeting or lamin A/C-targeting siRNA molecules, respectively. The results from these lines of study all suggest that redox signaling plays a central role in cytokine induction by DMXAA.http://www.sciencedirect.com/science/article/pii/S1476558610800776 |
| spellingShingle | Romy Brauer Liang-Chuan S. Wang See-Tarn Woon David J.A. Bridewell Kimiora Henare Dieter Malinger Brian D. Palmer Stefanie N. Vogel Claudine Kieda Sofian M. Tijono Lai-Ming Ching Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of Action Neoplasia: An International Journal for Oncology Research |
| title | Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of Action |
| title_full | Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of Action |
| title_fullStr | Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of Action |
| title_full_unstemmed | Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of Action |
| title_short | Labeling of Oxidizable Proteins with a Photoactivatable Analog of the Antitumor Agent DMXAA: Evidence for Redox Signaling in Its Mode of Action |
| title_sort | labeling of oxidizable proteins with a photoactivatable analog of the antitumor agent dmxaa evidence for redox signaling in its mode of action |
| url | http://www.sciencedirect.com/science/article/pii/S1476558610800776 |
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