Abstract 1852: Protein profiling during different stages of lung tumorigenesis induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in the A/J mouse model and the impact of chemopreventive agents
Authors: Bortner James D. , Das Arunangshu , Stanley Anne , Stanley Bruce , Aliaga Cesar , Cooper Timothy , Skibinski Christine , El-Bayoumy Karam 
Published: August 2014

In our previous proteomic study aimed at developing biomarkers for early detection of lung cancer, we identified several proteins that were differentially expressed in lung adenocarcinoma in A/J mice treated with the tobacco-specific nitrosamine NNK and were further modulated by the synthetic selenium chemopreventive agent 1,4-phenylenebis(methylene)selenocyanate (p-XSC). Unfortunately, many potential biomarkers described in previous reports, including our own, are based on comparative analysis between normal and end-stage lung tumors, and thus, can be considered as a consequence of tumor development rather than a cause of cancer induction. Therefore, in the present study we determined the effect of p-XSC and the naturally occurring chemopreventive agent myo-inositol (MI) on protein profiling during different stages of NNK-induced lung tumorigenesis; we used the proteomic approach isobaric Tags for Relative and Absolute Quantitation (iTRAQ). Four groups of mice were included in this study: vehicle, NNK, NNK+p-XSC, and NNK+MI; ten mice were sacrificed at 3 and 9 weeks after the first dose of NNK and additional mice will be sacrificed after 18, 24, and 31 weeks. Histopathology of the lungs showed airway and alveolar epithelial hyperplasia in all groups after 3 and 9 weeks, as well as airway epithelial atypia; however, no clear difference was observed. Equal amounts of total lung protein from five-mice/group were pooled and labeled with one of the isobaric tags 113-119, or 121. Mass spectrometry analysis led to identification of 790 proteins. After 3 and 9 weeks, 11 and 13 proteins, respectively, were significantly modulated in the lungs of mice treated with NNK vs. vehicle-treated mice; of these proteins, filamin-A was down-regulated, while clathrin heavy chain 1 was up-regulated after 3 and 9 weeks in NNK-treated mice; administration of either p-XSC or MI in NNK-treated mice had no apparent affect. After 3 weeks, the 60S ribosomal protein L3 was significantly down-regulated in mice treated with NNK alone, but was up-regulated in mice treated with NNK+p-XSC or MI; after 9 weeks no apparent change was associated with this protein for all groups. After 3 and 9 weeks, using immunoblot analysis, NNK had no apparent effect on 14-3-3 σ protein expression; however, it was increased in lungs of mice treated with NNK+p-XSC and not NNK+MI. The results of this study show that certain proteins in the lung are modulated after 3 and 9 weeks of NNK administration; the impact of p-XSC on NNK-induced protein alteration appears to vary from that observed with MI. Further validation of the proteins identified in this study and proteomic analysis after 18, 24, and 31 weeks of the bioassay will provide better insight into the selection of candidate protein biomarkers for lung cancer development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1852. doi:10.1158/1538-7445.AM2011-1852