ToxSci Advance Access published online on April 13, 2007
Toxicological Sciences, doi:10.1093/toxsci/kfm087
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Cyclophosphamide-Induced Apoptosis in COV434 Human Granulosa Cells Involves Oxidative Stress and Glutathione Depletion
,
,
* Dept. of Community and Environmental Medicine, University of California, Irvine CA 92617
Dept. of Medicine, University of California, Irvine CA 92617
Dept. of Developmental and Cell Biology, University of California, Irvine CA 92617
Dept. of Environmental Health, School of Public Health, Shanghai Jiaotong University, Shanghai, China
Corresponding Author: Dr. Ulrike Luderer, Center for Occupational and Environmental Health, 5201 California Avenue, Suite 100, Irvine, CA 92617, Tel: 949-824-8641, Fax: 949-824-2345, Email: uluderer{at}uci.edu
Received November 2, 2006; revision received April 7, 2007; accepted April 10, 2007
 |
Abstract |
|---|
The anti-cancer drug cyclophosphamide induces granulosa cell apoptosis and is detoxified by glutathione (GSH) conjugation. We previously showed that both cyclophosphamide treatment and GSH depletion induced granulosa cell apoptosis in rats, but the role of GSH in apoptosis in human ovarian cells has not been studied. Using the COV434 human granulosa cell line, we tested the hypotheses that 1) GSH depletion or treatment with 4-hydroperoxycyclophosphamide (4HC), a preactivated form of cyclophosphamide, induce apoptosis, 2) GSH depletion potentiates 4HC-induced apoptosis, 3) 4HC-induced apoptosis is mediated by GSH depletion and oxidative stress. Cells were treated with buthionine sulfoximine (BSO), a specific inhibitor of GSH synthesis, with or without follicle stimulating hormone (FSH) or serum. A significant increase in the number of apoptotic cells, assessed by TUNEL and Hoechst 33342 staining, occurred with BSO treatment. Treatment with 4HC dose-dependently induced apoptosis by TUNEL, Hoechst staining, and caspase 3 activation. Treatment with 4HC caused an increase in ROS generation, measured by dichlorofluorescein fluorescence, oxidative DNA damage, measured by 8-hydroxyguanosine immunostaining, and an oxidation of the redox potential for the oxidized glutathione/reduced glutathione (GSSG/2rGSH) couple. Total intracellular glutathione declined after 4HC treatment, preceding the onset of cell death. Treatment with antioxidants inhibited 4HC-induced apoptosis. Combined treatment with BSO and 4HC caused greater induction of apoptosis than either treatment alone. These findings are consistent with roles for oxidative stress and glutathione depletion in mediating the induction of apoptosis in COV434 cells by cyclophosphamide.
Key Words: granulosa cell; glutathione; cyclophosphamide; apoptosis; 4-hydroperoxycyclophosphamide; oxidative stress; ovary; antioxidant.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  Y. D. Hoang, B. N. Nakamura, and U. Luderer Follicle-Stimulating Hormone and Estradiol Interact to Stimulate Glutathione Synthesis in Rat Ovarian Follicles and Granulosa Cells Biol Reprod, October 1, 2009; 81(4): 636 - 646. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-Y. Choi, W. E. Barlow, K. S. Albain, C.-C. Hong, J. G. Blanco, R. B. Livingston, W. Davis, J. M. Rae, I-T. Yeh, L. F. Hutchins, et al. Nitric Oxide Synthase Variants and Disease-Free Survival among Treated and Untreated Breast Cancer Patients in a Southwest Oncology Group Clinical Trial Clin. Cancer Res., August 15, 2009; 15(16): 5258 - 5266. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Ortega-Camarillo, A. Gonzalez-Gonzalez, M. Vergara-Onofre, E. Gonzalez-Padilla, A. Avalos-Rodriguez, M. E Gutierrez-Rodriguez, L. Arriaga-Pizano, M. Cruz, L. A. Baiza-Gutman, and M. Diaz-Flores Changes in the glucose-6-phosphate dehydrogenase activity in granulosa cells during follicular atresia in ewes Reproduction, June 1, 2009; 137(6): 979 - 986. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. M. Cortes-Wanstreet, E. Giedzinski, C. L. Limoli, and U. Luderer Overexpression of glutamate-cysteine ligase protects human COV434 granulosa tumour cells against oxidative and {gamma}-radiation-induced cell death Mutagenesis, May 1, 2009; 24(3): 211 - 224. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Maman, K. Prokopis, J. Levron, A. Carmely, J. Dor, and D. Meirow Does controlled ovarian stimulation prior to chemotherapy increase primordial follicle loss and diminish ovarian reserve? An animal study Hum. Reprod., January 1, 2009; 24(1): 206 - 210. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Raffaghello, C. Lee, F. M. Safdie, M. Wei, F. Madia, G. Bianchi, and V. D. Longo From the Cover: Reactive Oxygen Species Special Feature: Starvation-dependent differential stress resistance protects normal but not cancer cells against high-dose chemotherapy PNAS, June 17, 2008; 105(24): 8215 - 8220. [Abstract] [Full Text] [PDF]
|
|