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Effect of Ionizing Radiation on Rat Lung Tissue: A Proteomic Analysis
I. J. Radiation Oncology d Biology d Physics
S618
Volume 69, Number 3, Supplement, 2007
Conclusions: Our study indicated that CD/UPRT expression, and to a less extent CD expression, significantly enhanced the efficacy of combined 5-FC and radiation treatment in R3327 prostate cancer cells. This suggests that CD/UPRT/5-FC strategy may have potential as an adjuvant to improve the outcome of radiotherapy. Author Disclosure: L. Xing, None; X. Deng, None; T.P.F. Gade, None; C.C. Ling, None; J.A. Koutcher, None; G.C. Li, None.
2757
Effect of Ionizing Radiation on Rat Lung Tissue: A Proteomic Analysis
P. R. Graves1, H. Sankala1, Z. Vujaskovic2 1
Virginia Commonwealth University, Richmond, VA, 2Duke University Medical School, Durham, NC
Purpose/Objective(s): The objective of this study is to examine the short and long-term effects of radiation on the rat lung proteome. Our goal is to determine proteomic changes associated with radiation-induced lung injury in order to identify early predictors of radiation response and potential therapeutic targets. Materials/Methods: Fischer 344 rats were irradiated with 150 kV x-rays at a single dose of 28 Gy delivered to the right hemithorax. Control animals were sham irradiated. Rats were sacrificed and lung tissue harvested post radiation at the following time points: 1, 3, 7, 10 days, 2, 4, 6, 8,12,14, and 20 wks. As an initial analysis of these complex samples, total lung cell lysates from each time point were resolved by one-dimensional gel electrophoresis and proteins visualized by silver staining. The proteins that showed differential levels of silver staining were excised and identified by mass spectrometry. Results: Significant changes in protein expression were observed at different times post irradiation (Fig. 1). The most significant ones are listed in Table 1. The process of validation of identified proteins by Western blotting is underway. Furthermore, a comprehensive analysis of irradiated lung tissue using PF 2D, a non-gel based method for 2-dimensional fractionation of complex protein samples is being conducted. Conclusions: Proteins upregulated after radiation are most commonly associated with cellular proliferation and adhesion, hypoxia, oxidative stress, and hemodynamic changes. In comparison, preliminary results show two antioxidants with important roles in pulmonary protection against oxidative stress, glutathione-s-transferase and peroxiredoxin-2, were downregulated post-radiation.
Table 1 Proteins Down-regulated by radiation 1. Filamin 2. Alpha-1 proteinase inhibitor III 3. 14-3-3 4. Carbonic Anhydrase and Glutathione-S-transferase 5. Rho-GDI alpha 6. Peroxiredoxin-2 Proteins Up-regulated by radition 7. Xanthine dehyrogenase/oxidase 8. Alpha spectrin-2 (Fodrin) 9. Protein disulfide isomerase 10. Cyclophilin B 11. Annexin II
Author Disclosure: P.R. Graves, None; H. Sankala, None; Z. Vujaskovic, None.
2758
Detection of Circulating Tumor Cells in the Peripheral Blood of Patients With Androgen-Independent, Advanced or Metastatic Prostate Cancer
D. T. Marshall, M. C. Mitas, U. B. Chaudhary, S. Gattoni-Celli Medical University of South Carolina, Charleston, SC Purpose/Objective(s): Accurate biochemical monitoring of late stage, androgen-independent prostate cancer (AIPC) is difficult and the correlation of clinical symptoms and disease progression with blood levels of prostate-specific antigen (PSA) is often poor. An increasing body of experimental evidence indicates that several types of cancer ‘‘leak’’ tumor cells into the bloodstream. The presence of circulating tumor cells (CTC) has been observed in colorectal and prostate cancer. Furthermore, CTC have been associated with poor prognosis in patients with metastatic breast cancer. This suggests that the monitoring of CTC could provide
S618
Volume 69, Number 3, Supplement, 2007
Conclusions: Our study indicated that CD/UPRT expression, and to a less extent CD expression, significantly enhanced the efficacy of combined 5-FC and radiation treatment in R3327 prostate cancer cells. This suggests that CD/UPRT/5-FC strategy may have potential as an adjuvant to improve the outcome of radiotherapy. Author Disclosure: L. Xing, None; X. Deng, None; T.P.F. Gade, None; C.C. Ling, None; J.A. Koutcher, None; G.C. Li, None.
2757
Effect of Ionizing Radiation on Rat Lung Tissue: A Proteomic Analysis
P. R. Graves1, H. Sankala1, Z. Vujaskovic2 1
Virginia Commonwealth University, Richmond, VA, 2Duke University Medical School, Durham, NC
Purpose/Objective(s): The objective of this study is to examine the short and long-term effects of radiation on the rat lung proteome. Our goal is to determine proteomic changes associated with radiation-induced lung injury in order to identify early predictors of radiation response and potential therapeutic targets. Materials/Methods: Fischer 344 rats were irradiated with 150 kV x-rays at a single dose of 28 Gy delivered to the right hemithorax. Control animals were sham irradiated. Rats were sacrificed and lung tissue harvested post radiation at the following time points: 1, 3, 7, 10 days, 2, 4, 6, 8,12,14, and 20 wks. As an initial analysis of these complex samples, total lung cell lysates from each time point were resolved by one-dimensional gel electrophoresis and proteins visualized by silver staining. The proteins that showed differential levels of silver staining were excised and identified by mass spectrometry. Results: Significant changes in protein expression were observed at different times post irradiation (Fig. 1). The most significant ones are listed in Table 1. The process of validation of identified proteins by Western blotting is underway. Furthermore, a comprehensive analysis of irradiated lung tissue using PF 2D, a non-gel based method for 2-dimensional fractionation of complex protein samples is being conducted. Conclusions: Proteins upregulated after radiation are most commonly associated with cellular proliferation and adhesion, hypoxia, oxidative stress, and hemodynamic changes. In comparison, preliminary results show two antioxidants with important roles in pulmonary protection against oxidative stress, glutathione-s-transferase and peroxiredoxin-2, were downregulated post-radiation.
Table 1 Proteins Down-regulated by radiation 1. Filamin 2. Alpha-1 proteinase inhibitor III 3. 14-3-3 4. Carbonic Anhydrase and Glutathione-S-transferase 5. Rho-GDI alpha 6. Peroxiredoxin-2 Proteins Up-regulated by radition 7. Xanthine dehyrogenase/oxidase 8. Alpha spectrin-2 (Fodrin) 9. Protein disulfide isomerase 10. Cyclophilin B 11. Annexin II
Author Disclosure: P.R. Graves, None; H. Sankala, None; Z. Vujaskovic, None.
2758
Detection of Circulating Tumor Cells in the Peripheral Blood of Patients With Androgen-Independent, Advanced or Metastatic Prostate Cancer
D. T. Marshall, M. C. Mitas, U. B. Chaudhary, S. Gattoni-Celli Medical University of South Carolina, Charleston, SC Purpose/Objective(s): Accurate biochemical monitoring of late stage, androgen-independent prostate cancer (AIPC) is difficult and the correlation of clinical symptoms and disease progression with blood levels of prostate-specific antigen (PSA) is often poor. An increasing body of experimental evidence indicates that several types of cancer ‘‘leak’’ tumor cells into the bloodstream. The presence of circulating tumor cells (CTC) has been observed in colorectal and prostate cancer. Furthermore, CTC have been associated with poor prognosis in patients with metastatic breast cancer. This suggests that the monitoring of CTC could provide