Comparative proteomic analysis of radiation-induced changes in mouse lung: fibrosis-sensitive and -resistant strains.
To determine whether comparative proteomics could detect differential protein expression after lung irradiation in two mouse strains with different radiation responses, lung proteins were subjected to two-dimensional orthogonal liquid-phase separations, with chromatofocusing in the first dimension and nonporous silica reverse-phase high-performance liquid chromatography (NPS-RP-HPLC) in the second. Five weeks after 12 Gy whole-lung irradiation, 15 and 31 proteins had significantly altered expression levels in C3H/HeJ (less likely to develop lung fibrosis) and C57BL/6J mice (more likely to develop lung fibrosis), respectively. These proteins were analyzed by HPLC-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) and identified by matching sequences in a peptide database. The proteins are associated with redox, energy consumption, glycolysis, or chromatin/ RNA structure formation. Five of the six redox-related proteins, including superoxide dismutase 1 (SOD1), cytochrome c oxidase, glutamate dehydrogenase, biliverdin reductase, peroxiredoxin and carbonyl reductase, were down-regulated in the irradiated C57BL/6J mice, whereas SOD1, sulfurtransferase and carbonyl reductase increased in the irradiated C3H/ HeJ mice. Thus decreased antioxidant proteins in the irradiated C57BL/6J mice may be correlated with increased early lung toxicity. Changes in SOD1 and 8-hydroxydeoxy-guanosine (8-OHdG, an oxidative stress marker) were further confirmed by immunohistochemistry and/or Western blot analysis. These data suggest that a proteomics approach has the potential to detect protein changes relevant to early lung toxicity after irradiation.
Ao
Xiaoping
X
Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Lubman
David M
DM
Davis
Mary A
MA
Xing
Xianying
X
Kong
Feng-Ming
FM
Lawrence
Theodore S
TS
Zhang
Ming
M
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000032:analysis; D008168:Lung / Q000737:chemistry / Q000528:radiation effects*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D040901:Proteomics; D011658:Pulmonary Fibrosis / Q000209:etiology / Q000378:metabolism*; D011836:Radiation Tolerance; D013482:Superoxide Dismutase / Q000032:analysis; D000072105:Superoxide Dismutase-1
D000080242:8-Hydroxy-2'-Deoxyguanosine; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D003849:Deoxyguanosine