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Urethane increases reactive oxygen species and activates extracellular signal-regulated kinase in RAW 264.7 macrophages and A549 lung epithelial cells

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Abstract

Urethane, which is used as an anesthetic for animal experiments, causes inflammation and cancer in the lung. BALB/c mice received 1 mg/g of urethane once a week for four consecutive weeks via intraperitoneal injections developed interstitial infiltration of inflammatory cells and tumors in the lung. However, the intracellular signaling events which urethane causes inflammation and cancer are largely unknown. Here we show that urethane caused overproduction of reactive oxygen species (ROS) in RAW 264.7 macrophages and A549 lung epithelial cells. Pretreatment of these cells with the antioxidant N-acetylcysteine attenuated the urethane-induced ROS production. Urethane increased heme oxygenase-1 expression to protect these cells from cytotoxicity caused by overproduced ROS. In addition, urethane activated extracellular signal-regulated kinase (ERK) in both cell types. Overall, our data imply that urethane stimulates ROS production and ERK activation in macrophages and lung epithelial cells, and the overproduced ROS and activated ERK may promote tumor formation in the lung.

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References

  • Allavena, P., C. Garlanda, M.G. Borrello, A. Sica, and A. Mantovani. 2008. Pathways connecting inflammation and cancer. Current Opinion in Genetics & Development 18: 3–10.

    Article  CAS  Google Scholar 

  • Azad, N., A.K. Iyer, A. Manosroi, L. Wang, and Y. Rojanasakul. 2008a. Superoxide-mediated proteasomal degradation of Bcl-2 determines cell susceptibility to Cr(VI)-induced apoptosis. Carcinogenesis 29: 1538–1545.

    Article  PubMed  CAS  Google Scholar 

  • Azad, N., Y. Rojanasakul, and V. Vallyathan. 2008b. Inflammation and lung cancer: roles of reactive oxygen/nitrogen species. Journal of Toxicology and Environmental Health Part B Critical Reviews. 11: 1–15.

    Article  CAS  Google Scholar 

  • Balansky, R.M., and S. De Flora. 1998. Chemoprevention by N-acetylcysteine of urethane-induced clastogenicity and lung tumors in mice. International Journal of Cancer 77: 302–305.

    Article  CAS  Google Scholar 

  • Chan, D.W., V.W. Liu, G.S. Tsao, K.M. Yao, T. Furukawa, K.K. Chan, and H.Y. Ngan. 2008. Loss of MKP3 mediated by oxidative stress enhances tumorigenicity and chemoresistance of ovarian cancer cells. Carcinogenesis 29: 1742–1750.

    Article  PubMed  CAS  Google Scholar 

  • Cooper, K.L., K.J. Liu, and L.G. Hudson. 2009. Enhanced ROS production and redox signaling with combined arsenite and UVA exposure: contribution of NADPH oxidase. Free Radical Biology and Medicine 47: 381–388.

    Article  PubMed  CAS  Google Scholar 

  • Coussens, L.M., and Z. Werb. 2002. Inflammation and cancer. Nature 420: 860–867.

    Article  PubMed  CAS  Google Scholar 

  • De Flora, S., M. Astengo, D. Serra, and C. Bennicelli. 1986. Inhibition of urethan-induced lung tumors in mice by dietary N-acetylcysteine. Cancer Letters 32: 235–241.

    Article  PubMed  Google Scholar 

  • Doris, K., S.P. Karabela, C.A. Kairi, D.C. Simoes, C. Roussos, S.G. Zakynthinos, I. Kalomenidis, T.S. Blackwell, and G.T. Stathopoulos. 2010. Allergic inflammation does not impact chemical-induced carcinogenesis in the lungs of mice. Respiratory Research 11: 118.

    Article  PubMed  Google Scholar 

  • Faux, S.P., T. Tai, D. Thorne, Y. Xu, D. Breheny, and M. Gaca. 2009. The role of oxidative stress in the biological responses of lung epithelial cells to cigarette smoke. Biomarkers 14(Suppl 1): 90–96.

    Article  PubMed  CAS  Google Scholar 

  • Hamilton Jr, R.F., S.A. Thakur, and A. Holian. 2008. Silica binding and toxicity in alveolar macrophages. Free Radical Biology and Medicine 44: 1246–1258.

    Article  PubMed  CAS  Google Scholar 

  • Hanausek, M., Z. Walaszek, A. Viaje, M. LaBate, E. Spears, D. Farrell, R. Henrich, A. Tveit, E.F. Walborg Jr, and T.J. Slaga. 2004. Exposure of mouse skin to organic peroxides: subchronic effects related to carcinogenic potential. Carcinogenesis 25: 431–437.

    Article  PubMed  CAS  Google Scholar 

  • Karoor, V., M. Le, D. Merrick, E.C. Dempsey, and Y.E. Miller. 2010. Vascular endothelial growth factor receptor 2-targeted chemoprevention of murine lung tumors. Cancer Prevention Research 3: 1141–1147.

    Article  PubMed  CAS  Google Scholar 

  • Kim, C., J.S. Jang, M.R. Cho, S.R. Agarawal, and Y.N. Cha. 2010. Taurine chloramine induces heme oxygenase-1 expression via Nrf2 activation in murine macrophages. International Immunopharmacology 10: 440–446.

    Article  PubMed  CAS  Google Scholar 

  • Kim, J.W., and C. Kim. 2005. Inhibition of LPS-induced NO production by taurine chloramine in macrophages is mediated through Ras-ERK-NF-kappaB. Biochemical Pharmacology 70: 1352–1360.

    Article  PubMed  CAS  Google Scholar 

  • Liu, L.Z., Y. Jiang, R.L. Carpenter, Y. Jing, S.C. Peiper, and B.H. Jiang. 2011. Role and mechanism of arsenic in regulating angiogenesis. PLoS ONE 6: e20858.

    Article  PubMed  CAS  Google Scholar 

  • McCubrey, J.A., M.M. Lahair, and R.A. Franklin. 2006. Reactive oxygen species-induced activation of the MAP kinase signaling pathways. Antioxidants & Redox Signaling 8: 1775–1789.

    Article  CAS  Google Scholar 

  • Nettleship, A., P. Henshaw, and H. Meyer. 1943. Induction of pulmonary tumors in mice with ethyl carbamate (urethane). Journal of the National Cancer Institute 4: 309–319.

    CAS  Google Scholar 

  • Nomura, T. 1975. Urethan (ethyl carbamate) as a cosolvent of drugs commonly used parenterally in humans. Cancer Research 35: 2895–2899.

    PubMed  CAS  Google Scholar 

  • Nomura, T. 1982. Parental exposure to X-rays and chemicals induces heritable tumours and anomalies in mice. Nature 296: 575–577.

    Article  PubMed  CAS  Google Scholar 

  • Nomura, T. 2008. Transgenerational effects from exposure to environmental toxic substances. Mutation Research 659: 185–193.

    Article  PubMed  CAS  Google Scholar 

  • O’Donnell, E.P., L.K. Zerbe, L.D. Dwyer-Nield, L.R. Kisley, and A.M. Malkinson. 2006. Quantitative analysis of early chemically-induced pulmonary lesions in mice of varying susceptibilities to lung tumorigenesis. Cancer Letters 241: 197–202.

    Article  PubMed  Google Scholar 

  • Orosz, Z., A. Csiszar, N. Labinskyy, K. Smith, P.M. Kaminski, P. Ferdinandy, M.S. Wolin, A. Rivera, and Z. Ungvari. 2007. Cigarette smoke-induced proinflammatory alterations in the endothelial phenotype: role of NAD(P)H oxidase activation. American Journal of Physiology Heart and Circulatory Physiology 292: H130–H139.

    Article  PubMed  CAS  Google Scholar 

  • Penza, M., C. Montani, M. Jeremic, G. Mazzoleni, W.L. Hsiao, M. Marra, H. Sharma, and D. Di Lorenzo. 2007. MAK-4 and -5 supplemented diet inhibit liver carcinogenesis in mice. BMC Complementary and Alternative Medicine 7: 19.

    Article  PubMed  Google Scholar 

  • Ramasamy, K., L.D. Dwyer-Nield, N.J. Serkova, K.M. Hasebroock, A. Tyagi, K. Raina, R.P. Singh, A.M. Malkinson, and R. Agarwal. 2010. Silibinin prevents lung tumorigenesis in wild-type but not in iNOS−/− mice: potential of real-time micro-CT in lung cancer chemoprevention studies. Clinical Cancer Research 17: 753–761.

    Article  PubMed  Google Scholar 

  • Rinehart, J., A.A. Adjei, P.M. Lorusso, D. Waterhouse, J.R. Hecht, R.B. Natale, O. Hamid, M. Varterasian, P. Asbury, E.P. Kaldjian, S. Gulyas, D.Y. Mitchell, R. Herrera, J.S. Sebolt-Leopold, and M.B. Meyer. 2004. Multicenter phase II study of the oral MEK inhibitor, CI-1040, in patients with advanced non-small-cell lung, breast, colon, and pancreatic cancer. Journal of Clinical Oncology 22: 4456–4462.

    Article  PubMed  CAS  Google Scholar 

  • Roomi, M.W., N.W. Roomi, T. Kalinovsky, M. Rath, and A. Niedzwiecki. 2009. Chemopreventive effect of a novel nutrient mixture on lung tumorigenesis induced by urethane in male A/J mice. Tumori 95: 508–513.

    PubMed  CAS  Google Scholar 

  • Rosin, F.C., J.F. Pedregosa, J.S. de Almeida, and V. Bueno. 2011. Identification of myeloid-derived suppressor cells and T regulatory cells in lung microenvironment after urethane-induced lung tumor. International Immunopharmacology 11: 873–878.

    Article  PubMed  CAS  Google Scholar 

  • Salinas, N.R., C.T. Lopes, P.V. Palma, C.T. Oshima, and V. Bueno. 2009. Lung tumor development in the presence of sphingosine 1-phosphate agonist FTY720. Pathology & Oncology Research 15: 549–554.

    Article  CAS  Google Scholar 

  • Saunders, J.A., L.C. Rogers, C. Klomsiri, L.B. Poole, and L.W. Daniel. 2010. Reactive oxygen species mediate lysophosphatidic acid induced signaling in ovarian cancer cells. Free Radical Biology and Medicine 49: 2058–2067.

    Article  PubMed  CAS  Google Scholar 

  • Singh, R.P., G. Deep, M. Chittezhath, M. Kaur, L.D. Dwyer-Nield, A.M. Malkinson, and R. Agarwal. 2006. Effect of silibinin on the growth and progression of primary lung tumors in mice. Journal of the National Cancer Institute 98: 846–855.

    Article  PubMed  CAS  Google Scholar 

  • Stathopoulos, G.T., T.P. Sherrill, D.S. Cheng, R.M. Scoggins, W. Han, V.V. Polosukhin, L. Connelly, F.E. Yull, B. Fingleton, and T.S. Blackwell. 2007. Epithelial NF-kappaB activation promotes urethane-induced lung carcinogenesis. Proceedings of the National Academy of Sciences of the United States of America 104: 18514–18519.

    Article  PubMed  CAS  Google Scholar 

  • Torres, M., and H.J. Forman. 2003. Redox signaling and the MAP kinase pathways. BioFactors 17: 287–296.

    Article  PubMed  CAS  Google Scholar 

  • Walser, T., X. Cui, J. Yanagawa, J.M. Lee, E. Heinrich, G. Lee, S. Sharma, and S.M. Dubinett. 2008. Smoking and lung cancer: the role of inflammation. Proceedings of the American Thoracic Society 5: 811–815.

    Article  PubMed  Google Scholar 

  • Yano, T., S. Yajima, K. Hagiwara, I. Kumadaki, Y. Yano, S. Otani, M. Uchida, and T. Ichikawa. 2000. Vitamin E inhibits cell proliferation and the activation of extracellular signal-regulated kinase during the promotion phase of lung tumorigenesis irrespective of antioxidative effect. Carcinogenesis 21: 2129–2133.

    Article  PubMed  CAS  Google Scholar 

  • Yano, T., Y. Yano, Y. Nagashima, M. Yuasa, S. Yajima, S. Horikawa, K. Hagiwara, M. Kishimoto, T. Ichikawa, and S. Otani. 1999. Activation of extracellular signal-regulated kinase in lung tissues of mice treated with carcinogen. Life Sciences 64: 229–236.

    Article  PubMed  CAS  Google Scholar 

  • Yao, H., I. Edirisinghe, S. Rajendrasozhan, S.R. Yang, S. Caito, D. Adenuga, and I. Rahman. 2008. Cigarette smoke-mediated inflammatory and oxidative responses are strain-dependent in mice. American Journal of Physiology Lung Cellular and Molecular Physiology 294: L1174–L1186.

    Article  PubMed  CAS  Google Scholar 

  • Zaynagetdinov, R., T.P. Sherrill, V.V. Polosukhin, W. Han, J.A. Ausborn, A.G. McLoed, F.B. McMahon, L.A. Gleaves, A.L. Degryse, G.T. Stathopoulos, F.E. Yull, and T.S. Blackwell. 2011. A critical role for macrophages in promotion of urethane-induced lung carcinogenesis. Journal of Immunology 187: 5703–5711.

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by Basic Science Research Program through the NRF of Korea (2009-0075348) and by a research grant from Inha University.

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  1. Laboratory for Leukocyte Signaling Research, Department of Pharmacology & Toxicology, Inha University School of Medicine, 7-206 3rd St, Shinheung-dong, Jung-gu, Incheon, 400-712, South Korea

    So-Hee Chun, Young-Nam Cha & Chaekyun Kim

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  1. So-Hee Chun
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Correspondence to Chaekyun Kim.

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Chun, SH., Cha, YN. & Kim, C. Urethane increases reactive oxygen species and activates extracellular signal-regulated kinase in RAW 264.7 macrophages and A549 lung epithelial cells. Arch. Pharm. Res. 36, 775–782 (2013). https://doi.org/10.1007/s12272-013-0104-8

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