In vitro Cytotoxic and Genotoxic Effects of Arsenic Trioxide on Human Keratinocytes
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.
[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.
[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.
@article{"International Journal of Medical, Medicine and Health Sciences:69179", author = "H. Bouaziz and M. Sefi and J. de Lapuente and M. Borras and N. Zeghal", title = "In vitro Cytotoxic and Genotoxic Effects of Arsenic Trioxide on Human Keratinocytes", abstract = "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 = "Arsenic trioxide, cytotoxixity, genotoxicity, HaCaT.", volume = "9", number = "3", pages = "231-4", }
