Abstract
While Cr (VI)‐containing compounds are well established carcinogens, the mechanisms of their action remain to be investigated. In this study we show that Cr (VI) causes increased tyrosine phosphorylation in human lung epithelial A549 cells in a time‐dependent manner. N‐acetyl‐cysteine (NAC), a general antioxidant, inhibited Cr (VI)‐induced tyrosine phosphorylation. Catalase, a scavenger of H2O2, sodium formate and aspirin, scavengers of hydroxyl radical (•OH), also inhibited the increased tyrosine phosphorylation induced by Cr (VI). SOD, an inhibitor of superoxide radical (O2 • −), caused less inhibition. ESR study shows that incubation of Cr (VI) with the A549 cells generates •OH radical. The generation of radical was decreased by addition of catalase and sodium formate, while SOD did not have any inhibitory effect. Oxygen consumption measurements show that addition of f Cr (VI) to A549 cells resulted in enhanced molecular oxygen consumption. These results indicate that Cr (VI) can induce an increase in tyrosine phosphorylation. H2O2 and •OH radicals generated during the process are responsible for the increased tyrosine phosphorylation induced by Cr (VI).
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De Flora S, Bagnasco M, Serra D, Zanacchi P: Genotoxicity of chromium compounds: a review. Mutat Res 238: 99–172, 1990
Shi X, Chiu A, Chen CT, Halliwell B, Castranova V, Vallyathan V: Reduction of chromium (VI) and its relationship to carcinogenesis. J Toxicol Environ Health 2: 87–104, 1999
De Flora S, Wetterhahn KE: Mechanism of chromium metabolism and genotoxicity. Life Chem Rep 7: 169–244, 1989
Xu J, Wise JP, Patierno SR: DNA damage induced by carcinogenic lead chromate particles in cultured mammalian cells. Mutat Res 280: 129–136, 1992
Leonard S, Wang S, Zang L, Castranova V, Vallyathan V, Shi X: Role of molecular oxygen in the generation of hydroxyl and superoxide anion radicals during enzymatic Cr (VI) reduction and its implication to Cr (VI)-induced carcinogenesis. J Environ Pathol Toxicol Oncol 19: 49–60, 2000
Xu J, Wise JP, Patierno SR: DNA damage induced by carcinogenic lead chromate particles in cultured mammalian cells. Mutat Res 280: 129–136, 1992
Kortenkamp A, Ozolins Z, Beyersmann D, O'Brien P: Generation of PM2 DNA breaks in the course of reduction of chromium (VI) by glutathione. Mutat Res 216: 19–26, 1989
Hayes RB: Review of occupational epidemiology of chromium chemicals and respiratory cancer. Sci Total Environ 71: 331–339, 1988
Langard S: One hundred years of chromium and cancer: A review of epidemiological evidence and selected case reports. Am J Ind Med 17: 189–215, 1990
Connett PH, Wetterhahn KE: Metabolism of the carcinogenic chromate by cellular constituents. Struct Bond 54: 93–124, 1983
Cohen MD, Kargacin B, Klein CB, Costa M: Mechanism of chromium carcinogenicity and toxicity. Crit Rev Toxicol 23: 255–281, 1983
Ye J, Wang S, Leonard SS, Sun Y, Butterworth, L, Antonini J, Ding M, Yongyut Rojanasakul, Vallyathan V, Castranova V, Shi X: Role of reactive oxygen species and p53 in chromium (VI)-induced apoptosis. J Biol Chem 274: 34974–34980, 1999
Ye J, Zhang X, Young HA, Mao Y, Shi X: Chromium (VI)-induced nuclear factor-κB activation in intact cells via free radical reactions. Carcinogenesis 16: 2401–2405, 1995
Chen F, Ding M, Lu Y, Leonard SS, Vallyathan V, Castranova V, Shi X: Participation of MAP kinase p38 and IκB kinase in chromium (VI)-induced NF-κB and AP-1 activation. J Environ Pathol Toxicol Oncol 19: 231–238, 2000
Wang S, Leonard SS, Ye J, Ding M, Shi X: The role of hydroxyl radical as a messenger in Cr (VI)-induced p53 activation. Am J Physiol 279: C868–C875, 2000
Chuang SM, Liou GY, Yang JL: Activation of JNK, p38, and ERK mitogen-activated protein kinase by chromium (VI) is mediated through oxidative stress but does not affect cytotoxicity. Carcinogenesis 21: 1491–1500, 2000
Chuang SM, Wang IC, Yang JL: Roles of JNK, p38 and ERK mitogenactivated protein kinase in the growth inhibition and apoptosis induced by cadmium. Carcinogenesis 21: 1423–1432, 2000
Gozin A, Franzini E, Andrieu V, Da Costa L, Rollet-Labelle E, Pasquier C: Reactive oxygen species activate focal adhesion kinase, paxillin and tyrosine phosphorylation in endothelial cells. Free Rad Biol Med 9: 1021–1032, 1998
Hunter T: The Croonian Lecture 1997. The phosphorylation of proteins on tyrosine: Its role in cell growth and disease. Phil Trans R Soc Lond B Biol Sci 353: 583–605, 1998
Jiang G, Hunter T: Receptor signaling: When dimerization is not enough. Curr Biol 9: R568–R571, 1999
Brown MT, Cooper JA: Regulation, substrates and functions of src. Biochim Biophys Acta 1287: 121–149, 1996
Glenney JR Jr: Tyrosine-phosphorylated proteins: Mediators of signal transduction from the tyrosine kinases. Biochim Biophys Acta 1134: 113–127, 1992
Kolibaba KS, Druker BJ: Protein tyrosine kinases and cancer. Biochim Biophys Acta. 1333: F217–F248, 1997
Biscardi JS, Tice DA, Parsons SJ: c-Src, receptor tyrosine kinases, and human cancer. Adv Cancer Res 76: 61–119, 1999
Hill CS, Treisman R: Transcriptional regulation by extracellular signals: Mechanisms and specificity. Cell 80: 199–211, 1995
Darnell JE Jr, Kerr IM, Stark GR: Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science 264: 1415–1421, 1994
Imbert V, Rupec RA, Livolsi A, Pahl HL, Traenckner EB, Mueller-Dieckmann C, Farahifar D, Rossi B, Auberger P, Baeuerle PA, Peyron JF: Tyrosine phosphorylation of IκB-alpha activates NF-κB without proteolytic degradation of IκB alpha. Cell 86: 787–798, 1996
Buettner GR: ESR parameters of spin adducts. Free Rad Biol Med 3: 259–303, 1987
Halliwell B, Gutteridge JMC: In: Free Radicals in Biology and Medicine. Oxford University Press, Oxford, UK, 2000, pp 1–540
Vallyathan V, Shi X: The role of free radicals in occupational and environmental lung diseases. Environ Health Perspect 105: 195–177, 2000
Pennisi E: Superoxides relay Ras protein's oncogenic message. Science 1567–1568, 1997
Buzard GS, Kasprzak KS: Possible role of nitroxide and redox cell signaling in metal-induced toxicity and carcinogenesis: A review. J Environ Pathol Toxicol Oncol 19: 179–199, 2000
Irani K, Xia Y, Zweier JL, Sollott SJ, Der CJ, Fearon ER, Sundaresan M, Finkel T, Goldschmidt-Clermont PJ: Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts. Science 275: 1649–1652, 1997
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Qian, Y., Jiang, B., Flynn, D.C. et al. Cr(VI) increases tyrosine phosphorylation through reactive oxygen species-mediated reactions. Mol Cell Biochem 222, 199–204 (2001). https://doi.org/10.1023/A:1017926531890
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DOI: https://doi.org/10.1023/A:1017926531890