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Mustard gas surrogate, 2-chloroethyl ethylsulfide (2-CEES), induces centrosome amplification and aneuploidy in human and mouse cells

2-CEES induces centrosome amplification and chromosome instability

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Abstract

Mustard gas is a simple molecule with a deadly past. First used as a chemical weapon in World War I, its simple formulation has raised concerns over its use by terrorist organizations and unstable governments. Mustard gas is a powerful vesicant and alkylating agent that causes painful blisters on epithelial surfaces and increases the incidence of cancer in those exposed. The mechanism of mustard gas toxicity and tumorigenesis is not well understood but is thought to be mediated by its ability to induce oxidative stress and DNA damage. Interestingly, several proteins that have been shown to either be targets of mustard gas or mediate mustard gas toxicity have also been shown to regulate centrosome duplication. Centrosomes are small nonmembrane-bound organelles that direct the segregation of chromosomes during mitosis through the formation of the bipolar mitotic spindle. Cells with more or less than two centrosomes during mitosis can segregate their chromosomes unequally, resulting in chromosome instability, a common phenotype of cancer cells. In our studies, we show that subtoxic levels of 2-chloroethyl ethylsulfide (2-CEES), a mustard gas analog, induce centrosome amplification and chromosome instability in cells, which may hasten the mutation rate necessary for tumorigenesis. These data may explain why those exposed to mustard gas exhibit higher incidences of cancer than unexposed individuals of the same cohort.

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Abbreviations

2-CEES:

2-Chloroethyl ethylsulfide

CIN:

Chromosome instability

MT:

Microtubule

PARP:

Poly(ADP-ribose) polymerase

PBS:

Phosphate-buffered saline

DMEM:

Dulbecco’s Modified Eagle Medium

TBS:

Tris-buffered saline

DAPI:

4′,6-Diamidino-2-phenylindole

ROS:

Reactive oxygen species

References

  • Aasted A, Darre E, Wulf HC. Mustard gas: clinical, toxicological, and mutagenic aspects based on modern experience. Ann Plast Surg. 1987;19:330–3.

    Article  CAS  PubMed  Google Scholar 

  • Altman SA, Randers L, Rao G. Comparison of trypan blue dye exclusion and fluorometric assays for mammalian cell viability determinations. Biotechnol Prog. 1993;9:671–4.

    Article  CAS  PubMed  Google Scholar 

  • Azizi F, Keshavarz A, Roshanzamir F, Nafarabadi M. Reproductive function in men following exposure to chemical warfare with sulphur mustard. Med War. 1995;11:34–44.

    Article  CAS  PubMed  Google Scholar 

  • Bennett RA, Izumi H, Fukasawa K. Induction of centrosome amplification and chromosome instability in p53-null cells by transient exposure to subtoxic levels of S-phase-targeting anticancer drugs. Oncogene. 2004;23:6823–9.

    Article  CAS  PubMed  Google Scholar 

  • Bhat KR, Benton BJ, Rosenthal DS, Smulson ME, Ray R. Role of poly(adp-ribose) polymerase (parp) in DNA repair in sulfur mustard-exposed normal human epidermal keratinocytes (nhek). J Appl Toxicol. 2000;20 Suppl 1:S13–7.

    CAS  PubMed  Google Scholar 

  • Chae S, Yun C, Um H, Lee JH, Cho H. Centrosome amplification and multinuclear phenotypes are induced by hydrogen peroxide. Exp Mol Med. 2005;37:482–7.

    Article  CAS  PubMed  Google Scholar 

  • Cook JR, Van Buskirk RG. A double-label technique that monitors sulfur mustard damage to nuclei and mitochondria of normal human epidermal keratinocytes in vitro. Toxicol Pathol. 1997;25:481–6.

    Article  CAS  PubMed  Google Scholar 

  • Dillman 3rd JF, Phillips CS, Dorsch LM, Croxton MD, Hege AI, Sylvester AJ, et al. Genomic analysis of rodent pulmonary tissue following bis-(2-chloroethyl) sulfide exposure. Chem Res Toxicol. 2005;18:28–34.

    Article  CAS  PubMed  Google Scholar 

  • Doi M, Hattori N, Yokoyama A, Onari Y, Kanehara M, Masuda K, et al. Effect of mustard gas exposure on incidence of lung cancer: a longitudinal study. Am J Epidemiol. 2011;173:659–66.

    Article  PubMed  Google Scholar 

  • Duchovic RJ, Vilensky JA. Mustard gas: its pre-World War I history. J Chem Educ. 2007;84:944–8.

    Article  CAS  Google Scholar 

  • Duelfer C. NPR. Syria’s chemical weapons include sarin, mustard gas. 2012; http://www.npr.org/2012/12/07/166755923/syrias-chemical-weapons-include-sarin-mustard-gas [31 May 2013].

  • Easton DF, Peto J, Doll R. Cancers of the respiratory tract in mustard gas workers. Br J Ind Med. 1988;45:652–9.

    CAS  PubMed Central  PubMed  Google Scholar 

  • Everley PA, Dillman 3rd JF. A large-scale quantitative proteomic approach to identifying sulfur mustard-induced protein phosphorylation cascades. Chem Res Toxicol. 2010;23:20–5.

    Article  CAS  PubMed  Google Scholar 

  • Fidder A, Moes GW, Sheffer AG, van der Schans GP, Baan RA, de Jong LP, et al. Synthesis, characterization, and quatitation of the major adducts formed between sulfur mustard and DNA of calf thymus and human blood. Chem Res Toxicol. 1994;7:199–204.

    Article  CAS  PubMed  Google Scholar 

  • Fukasawa K. Centrosome amplification, chromosome instability and cancer development. Cancer Lett. 2005;230:6–19.

    Article  CAS  PubMed  Google Scholar 

  • Fukasawa K, Choi T, Kuriyama R, Rulong S, Vande Woude GF. Abnormal centrosome amplification in the absence of p53. Science. 1996;271:1744–7.

    Article  CAS  PubMed  Google Scholar 

  • Gadsden-Gray J, Mukherjee S, Ogunkua O, Das SK. Induction of neuronal damage in guinea pig brain by intratracheal infusion of 2-chloroethyl ethyl sulfide, a mustard gas analog. J Biochem Mol Toxicol. 2012;26:23–30.

    Article  CAS  PubMed  Google Scholar 

  • Gholamrezanezhad A, Saghari M, Vakili A, Mirpour S, Farahani MH. Myocardial perfusion abnormalities in chemical warfare patients intoxicated with mustard gas. Int J Cardiovasc Imaging. 2007;23:197–205.

    Article  PubMed  Google Scholar 

  • Hay A. Effects on health of mustard gas. Nature. 1993;366:398.

    Article  CAS  PubMed  Google Scholar 

  • Hosseini-khalili A, Haines DD, Modirian E, Soroush M, Khateri S, Joshi R, et al. Mustard gas exposure and carcinogenesis of lung. Mutat Res. 2009;678:1–6.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hough A. The Telegraph. Libya: mustard gas ‘most potent chemical weapon’. 2011; http://www.telegraph.co.uk/news/worldnews/africaandindianocean/libya/8353787/Libya-mustard-gas-most-potent-chemical-weapon.html [31 May 2013].

  • Inturi S, Tewari-Singh N, Gu M, Shrotriya S, Gomez J, Agarwal C, et al. Mechanisms of sulfur mustard analog 2-chloroethyl ethyl sulfide-induced DNA damage in skin epidermal cells and fibroblasts. Free Radic Biol Med. 2011;51:2272–80.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Iravani S, Rahnavardi M, Gorouhi F, Gorouhi F. Repeated gastrointestinal malignancies in a victim of sulfur mustard gas attack. Indian J Gastroenterol. 2007;26:102.

    PubMed  Google Scholar 

  • Jowsey PA, Williams FM, Blain PG. DNA damage, signalling and repair after exposure of cells to the sulphur mustard analogue 2-chloroethyl ethyl sulphide. Toxicology. 2009;257:105–12.

    Article  CAS  PubMed  Google Scholar 

  • Jowsey PA, Williams FM, Blain PG. DNA damage responses in cells exposed to sulphur mustard. Toxicol Lett. 2012;209:1–10.

    Article  CAS  PubMed  Google Scholar 

  • Kanai M, Tong WM, Sugihara E, Wang ZQ, Fukasawa K, Miwa M. Involvement of poly(adp-ribose) polymerase 1 and poly(adp-ribosyl)ation in regulation of centrosome function. Mol Cell Biol. 2003;23:2451–62.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Liu JL, Ma HP, Lu XL, Sun SH, Guo X, Li FC. Nf-kappab induces abnormal centrosome amplification by upregulation of cdk2 in laryngeal squamous cell cancer. Int J Oncol. 2011;39:915–24.

    CAS  PubMed  Google Scholar 

  • Malhotra RC, Ganesan K, Sungedran K, Swamy RV. Chemistry and Toxicology of Sulphur Mustard - A Review. Def Sci J. 1999;49(2), 97--116.

  • Nigg EA, Raff JW. Centrioles, centrosomes, and cilia in health and disease. Cell. 2009;139:663–78.

    Article  CAS  PubMed  Google Scholar 

  • Nishimoto Y, Burrows B, Miyanishi M, Katsuta S, Shigenobu T, Kettel LJ. Chronic obstructive lung disease in japanese poison gas workers. Am Rev Respir Dis. 1970;102:173–9.

    CAS  PubMed  Google Scholar 

  • Nishimoto Y, Yamakido M, Shigenobu T, Onari K, Yukutake M. Long-term observation of poison gas workers with special reference to respiratory cancers. J UOEH. 1983;5(Suppl):89–94.

    PubMed  Google Scholar 

  • O'Brien J, Wilson I, Orton T, Pognan F. Investigation of the alamar blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem. 2000;267:5421–6.

    Article  PubMed  Google Scholar 

  • Pal A, Tewari-Singh N, Gu M, Agarwal C, Huang J, Day BJ, et al. Sulfur mustard analog induces oxidative stress and activates signaling cascades in the skin of skh-1 hairless mice. Free Radic Biol Med. 2009;47:1640–51.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Pihan GA, Purohit A, Wallace J, Knecht H, Woda B, Quesenberry P, et al. Centrosome defects and genetic instability in malignant tumors. Cancer Res. 1998;58:3974–85.

    CAS  PubMed  Google Scholar 

  • Qui M, Paromov VM, Yang H, Smith M, Stone WL. Inhibition of inducible nitric oxide synthase by a mustard gas analog in murine macrophages. BMC Cell Biol. 2006;7:39.

    Article  PubMed Central  PubMed  Google Scholar 

  • Rancourt RC, Veress LA, Guo X, Jones TN, Hendry-Hofer TB, White CW. Airway tissue factor-dependent coagulation activity in response to sulfur mustard analog 2-chloroethyl ethyl sulfide. Am J Physiol Lung Cell Mol Physiol. 2012;302:L82–92.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Rosenthal DS, Simbulan-Rosenthal CM, Iyer S, Smith WJ, Ray R, Smulson ME. Calmodulin, poly(adp-ribose)polymerase and p53 are targets for modulating the effects of sulfur mustard. J Appl Toxicol. 2000;20 Suppl 1:S43–9.

    CAS  PubMed  Google Scholar 

  • Ross WCJ. Biological alkylating agents. London: Butterworths; 1962.

    Google Scholar 

  • Ruff AL, Dillman 3rd JF. Sulfur mustard induced cytokine production and cell death: Investigating the potential roles of the p38, p53, and nf-kappab signaling pathways with rna interference. J Biochem Mol Toxicol. 2010;24:155–64.

    Article  CAS  PubMed  Google Scholar 

  • Scholz RO, Woods AC. Relapsing and chronic ocular lesions following mustard gas (dichloroethyl sulfide) burns. Arch Ophthalmol. 1947;37:139–48.

    Article  CAS  Google Scholar 

  • Sharma DR, Sunkaria A, Bal A, Bhutia YD, Vijayaraghavan R, Flora SJ, et al. Neurobehavioral impairments, generation of oxidative stress and release of pro-apoptotic factors after chronic exposure to sulphur mustard in mouse brain. Toxicol Appl Pharmacol. 2009;240:208–18.

    Article  CAS  PubMed  Google Scholar 

  • Shinmura K, Bennett RA, Tarapore P, Fukasawa K. Direct evidence for the role of centrosomally localized p53 in the regulation of centrosome duplication. Oncogene. 2007;26:2939–44.

    Article  CAS  PubMed  Google Scholar 

  • Smith KJ, Hurst CG, Moeller RB, Skelton HG, Sidell FR. Sulfur mustard: Its continuing threat as a chemical warfare agent, the cutaneous lesions induced, progress in understanding its mechanism of action, its long-term health effects, and new developments for protection and therapy. J Am Acad Dermatol. 1995;32:765–76.

    Article  CAS  PubMed  Google Scholar 

  • Stoppler H, Stoppler MC, Johnson E, Simbulan-Rosenthal CM, Smulson ME, Iyer S, et al. The e7 protein of human papillomavirus type 16 sensitizes primary human keratinocytes to apoptosis. Oncogene. 1998;17:1207–14.

    Article  CAS  PubMed  Google Scholar 

  • Takeshima Y, Inai K, Bennett WP, Metcalf RA, Welsh JA, Yonehara S, et al. P53 mutations in lung cancers from Japanese mustard gas workers. Carcinogenesis. 1994;15:2075–9.

    Article  CAS  PubMed  Google Scholar 

  • Tarapore P, Horn HF, Tokuyama Y, Fukasawa K. Direct regulation of the centrosome duplication cycle by the p53-p21Waf1/Cip1 pathway. Oncogene. 2001;20:3173–84.

    Article  CAS  PubMed  Google Scholar 

  • Tewari-Singh N, Agarwal C, Huang J, Day BJ, White CW, Agarwal R. Efficacy of glutathione in ameliorating sulfur mustard analog-induced toxicity in cultured skin epidermal cells and in skh-1 mouse skin in vivo. J Pharmacol Exp Ther. 2011;336:450–9.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Thomsen AB, Eriksen J, Smidt-Nielsen K. Chronic neuropathic symptoms after exposure to mustard gas: a long-term investigation. J Am Acad Dermatol. 1998;39:187–90.

    Article  CAS  PubMed  Google Scholar 

  • Tong WM, Yang YG, Cao WH, Galendo D, Frappart L, Shen Y, et al. Poly(adp-ribose) polymerase-1 plays a role in suppressing mammary tumourigenesis in mice. Oncogene. 2007;26:3857–67.

    Article  CAS  PubMed  Google Scholar 

  • Vogelstein B, Lane D, Levine AJ. Surfing the p53 network. Nature. 2000;408:307–10.

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Liu L, Montagna C, Ried T, Deng CX. Haploinsufficiency of parp1 accelerates brca1-associated centrosome amplification, telomere shortening, genetic instability, apoptosis, and embryonic lethality. Cell Death Differ. 2007;14:924–31.

    CAS  PubMed  Google Scholar 

  • Wang QQ, Begum RA, Day VW, Bowman-James K. Sulfur, oxygen, and nitrogen mustards: stability and reactivity. Org Biomol Chem. 2012;10:8786–93.

    Article  CAS  PubMed  Google Scholar 

  • Wheeler GP. Studies related to the mechanisms of action of cytotoxic alkylating agents: a review. Cancer Res. 1962;22:651–88.

  • Yamakido M, Ishioka S, Hiyama K, Maeda A. Former poison gas workers and cancer: Incidence and inhibition of tumor formation by treatment with biological response modifier n-cws. Environ Health Perspect. 1996;104 Suppl 3:485–8.

    Article  PubMed Central  PubMed  Google Scholar 

  • Zojaji R, Balali-Mood M, Mirzadeh M, Saffari A, Maleki M. Delayed head and neck complications of sulphur mustard poisoning in Iranian veterans. J Laryngol Otol. 2009;123:1150–4.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors would like to thank the University of Southern Indiana Endeavor! Awards committee, Summer Research Fellowship committee, and the STEM-Early Undergraduate Research Program for the financial support of these experiments. We would also like to thank the biology and chemistry departments for technical assistance and Dr. Carla Aldrich, Dr. Jonathan Stallings, and Dr. Elisia Tichy for the manuscript review.

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Authors and Affiliations

  1. Biology Department, University of Southern Indiana, Evansville, IN, 47712, USA

    Richard A. Bennett, Elizabeth Behrens & Christian Duncheon

  2. Department of Biological Sciences, University of Toledo, Toledo, OH, 43606, USA

    Ashtyn Zinn

  3. Air Force Research Laboratory, Wright-Patterson AFB, Dayton, OH, 45433-7913, USA

    Thomas J. Lamkin

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  1. Richard A. Bennett
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  2. Elizabeth Behrens
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  3. Ashtyn Zinn
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  4. Christian Duncheon
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  5. Thomas J. Lamkin
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Correspondence to Richard A. Bennett or Thomas J. Lamkin.

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Bennett, R.A., Behrens, E., Zinn, A. et al. Mustard gas surrogate, 2-chloroethyl ethylsulfide (2-CEES), induces centrosome amplification and aneuploidy in human and mouse cells. Cell Biol Toxicol 30, 195–205 (2014). https://doi.org/10.1007/s10565-014-9279-0

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