Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 31242
In vitro Cytotoxic and Genotoxic Effects of Arsenic Trioxide on Human Keratinocytes

Authors: H. Bouaziz, M. Sefi, J. de Lapuente, M. Borras, N. Zeghal


Although, arsenic trioxide has been the subject of toxicological research, in vitro cytotoxicity and genotoxicity studies using relevant cell models and uniform methodology are not well elucidated. Hence, the aim of the present study was to evaluate the cytotoxicity and genotoxicity induced by arsenic trioxide in human keratinocytes (HaCaT) using the MTT [3-(4, 5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide] and alkaline single cell gel electrophoresis (Comet) assays, respectively. Human keratinocytes were treated with different doses of arsenic trioxide for 4 h prior to cytogenetic assessment. Data obtained from the MTT assay indicated that arsenic trioxide significantly reduced the viability of HaCaT cells in a dose-dependent manner, showing an IC50 value of 34.18 ± 0.6 μM. Data generated from the comet assay also indicated a significant dose-dependent increase in DNA damage in HaCaT cells associated with arsenic trioxide exposure. We observed a significant increase in comet tail length and tail moment, showing an evidence of arsenic trioxide -induced genotoxic damage in HaCaT cells. This study confirms that the comet assay is a sensitive and effective method to detect DNA damage caused by arsenic.

Keywords: Genotoxicity, arsenic trioxide, cytotoxixity, HaCaT

Digital Object Identifier (DOI):

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1936


[1] NRC, “Arsenic in drinking water.” U.S. National Research Council, National Academy Press. Washington, DC, 1999.
[2] P. Rousselot, S. Labaume, JP. Marolleau, J. Larghero, MK. Noguera, JC. Brouet, and JP. Ermand, “Arsenic trioxide and melarsoprol induce apoptosis in plasma cell lines and in plasma cells from myeloma patients.” Cancer Res, vol. 59, pp. 1041–1048, 1999.
[3] K. Zhang, K. Ohnishi, Shigeno, S. Fujisawa, K. Naito, S. Nakamura, K. Takeshita, A. Akeshita, and R. Ohno, “The induction of apoptosis and cell cycle arrest by arsenic trioxide in lymphoid neoplasms.” Leukemia vol. 12, pp.1383–1391, 1998.
[4] IARC, “Monographs on the Evaluation of Carcinogenic Risks of Chemicals to Humans.” Supplement F. Overall Evaluation of Carcinogenicity. International Agency for Research on Cancer. World Health Organization. Lyon, France, pp 29–57, 1987.
[5] A. Aschengran, S. Zierler, and A. Cohen, “Quality of community drinking water and the occurrence of spontaneous abortion.” Arch Environ Health vol. 44, pp. 283–290, 1989.
[6] JC. Barrett, PW. Lamb, TC. Wang, and TC. Lee, “Mechanisms of arsenic-induced cell transformation.” Biol Trace Elem Res vol. 21, pp. 421–429, 1989.
[7] B. Saleha Banu, K. Danadevi, J. Kaiser, YR. Ahuja, K. Visweswara Rao, and M. Ishap, “In vivo genotoxic effect of arsenic trioxide in mice using comet assay.” Toxicol vol. 162, pp. 171–177, 2001.
[8] PB. Tchounwou, CG. Yedjou, D. Foxx, A. Ishaque, and E. Shen, “Lead– induced cytotoxicity and transcriptional activation of stress genes in human liver carcinoma cells.” Mol Cell Biochem vol. 255, pp. 161–170, 2004.
[9] Y. Jing, J. Dai, RM. Charmers-Redman, WG. Tatto, and S. Waxman, “Arsenic trioxide selectively induces acute promyelocytic leukemia cell apoptosis via a hydrogen peroxide dependent pathway.” Blood vol. 94, pp. 102–2111, 1999.
[10] P. Jai, G. Chen, X. Huang, X. Cai, J. Yang, L. Wang, Y. Zhou, Y. Shen, L. Zhou, Y. Yu, S. Chen, X. Zhang, and Z. Wang, “Arsenic trioxide induces multiple myeloma cell apoptosis via disruption of mitochondrial transmembrane potentials and activation of caspace-3.” Chin Med J vol. 114, pp. 19–24, 1999.
[11] SL. Soignet, SR. Frankel, D. Douer, MS. Tallman, H. Kantarjian, E. Calleja, RM. Stone, M. Kalaycio, DA. Scheinberg, P. Steinherz, EL. Sievers, S. Coutre´, S. Dahlberg, R. Ellison, and RP. Warrell, “United States multicenter study of arsenic trioxide in relapsed acute promyelocytic leukemia.” J Clin Oncol vol. 19, pp. 3852–3860, 2001.
[12] O. Ostling, KJ. Johanson, “Microelectrophoretic study of radiationinduced DNA damages in individual mammalian cells.” Biochem Biophys Res Commun vol. 123, pp. 291–298, 1984.
[13] E. Rojas, MC. Lopez, and M. Valverde, “Single gel electrophoresis assay: methodology and application." J Chromatogr vol. 722, pp. 225– 254, 1999.
[14] M. Jaiswal, G. Anuradha, N. Rajeswari, KN. Raj, N. Balakrishna, KV. Rao, IS. Prasand, SN. Jain, and YR. Ahuja, “Comet assay on cervical epithelial cells and leucocytes of patients with precancerous and cancerous lesions of the cervix.” Med Sci Res vol. 22, pp. 879–881, 1994.