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Effect of Light Irradiation and Sex Hormones on Jurkat T Cells: 17β-Estradiol but Not Testosterone Enhances UVA-Induced Cytotoxicity in Jurkat Lymphocytes
Department of Surgery, Division of Plastic Surgery, University of Mississippi Medical Center, 2500 North State Street, Jackson, Mississippi, 39216-4505, USA
Department of Medicine, Division of Rheumatology/Immunology, University of Mississippi Medical Center, 2500 North State Street, Jackson, Mississippi, 39216-4505, USA
Department of Chemistry, Jackson State University, Jackson, Mississippi 39217, USA
* Author to whom correspondence should be addressed.
Received: 15 November 2004 / Accepted: 6 February 2005 / Published: 30 April 2005
Abstract: In Eastern cultures, such as India, it is traditionally recommended that women but not men cover their heads while working in the scorching sun. The purpose of this pilot study was to determine whether there was any scientific basis for this cultural tradition. We examined the differential cytotoxic effects of ultraviolet A light (UVA) on an established T cell line treated with female and male sex hormones. CD4+ Jurkat T cells were plated in 96 well plates at 2 x 106 cells/ml and treated with 17β-estradiol (EST) or testosterone (TE). These cells were irradiated by UVA light with an irradiance of 170 J/cm2 for 15min at a distance of 6 cm from the surface of the 96-well plate. Controls included cells not treated with hormones or UVA. The effects of EST and TE were investigated between 1 and 20 ng/mL. Cytotoxicity by fluorescein-diacetate staining and COMET assay generating single strand DNA cleavage, tail length and tail moment measurements were examined. The effect of estrogen (5ng/mL) on apoptosis and its mediators was further studied using DNA laddering and western blotting for bcl-2 and p53. We found that EST alone, without UVA, enhanced Jurkat T cell survival. However, EST exhibited a dose-related cytotoxicity in the presence of UVA; up to 28% at 20 ng/ml. TE did not alter UVA-induced cytotoxicity. Since TE did not alter cell viability in the presence of UVA further damaging studies were not performed. COMET assay demonstrated the harmful effects of EST in the presence of UVA while EST without UVA had no significant effect on the nuclear damage. Apoptosis was not present as indicated by the absence of DNA laddering on agarose gel electrophoresis at 5ng/ml EST or TE ± UVA. Western blot showed that estrogen down regulated bcl-2 independently of UVA radiation while p53 was down regulated in the presence of UVA treatment. EST and TE have differential effects on UVA-induced cytotoxicity in Jurkat T-lymphocyte which suggested that women may be more susceptible to the harmful effects of solar irradiation than men.
Keywords: apoptosis; bcl-2; comet assay; cytotoxicity assay; estrogen; Jurkat T cells; p53; testosterone; ultraviolet light A
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Cohly, H.H.; Graham-Evans, B.; Ndebele, K.; Jenkins, J.K.; McMurray, R.; Yan, J.; Yu, H.; Angel, M.F. Effect of Light Irradiation and Sex Hormones on Jurkat T Cells: 17β-Estradiol but Not Testosterone Enhances UVA-Induced Cytotoxicity in Jurkat Lymphocytes. Int. J. Environ. Res. Public Health 2005, 2, 156-163.
Cohly HH, Graham-Evans B, Ndebele K, Jenkins JK, McMurray R, Yan J, Yu H, Angel MF. Effect of Light Irradiation and Sex Hormones on Jurkat T Cells: 17β-Estradiol but Not Testosterone Enhances UVA-Induced Cytotoxicity in Jurkat Lymphocytes. International Journal of Environmental Research and Public Health. 2005; 2(1):156-163.
Cohly, Hari H.; Graham-Evans, Barbara; Ndebele, Kenneth; Jenkins, John K.; McMurray, Robert; Yan, Jian; Yu, Hongtao; Angel, Michael F. 2005. "Effect of Light Irradiation and Sex Hormones on Jurkat T Cells: 17β-Estradiol but Not Testosterone Enhances UVA-Induced Cytotoxicity in Jurkat Lymphocytes." Int. J. Environ. Res. Public Health 2, no. 1: 156-163.