title: Glucocorticoid receptor knockdown decreases the antioxidant protection of B16 melanoma cells: an endocrine system-related mechanism that compromises metastatic cell resistance to vascular endothelium-induced tumor cytotoxicity.
abstract: We previously reported an interorgan system in which stress-related hormones (corticosterone and noradrenaline), interleukin-6, and glutathione (GSH) coordinately regulate metastatic growth of highly aggressive B16-F10 melanoma cells. Corticosterone, at levels measured in tumor-bearing mice, also induces apoptotic cell death in metastatic cells with low GSH content. In the present study we explored the potential role of glucocorticoids in the regulation of metastatic cell death/survival during the early stages of organ invasion. Glucocorticoid receptor (GCR) knockdown decreased the expression and activity of γ-glutamylcysteine synthetase (γ-GCS), the rate-limiting step in GSH synthesis, in metastatic cells in vivo independent of the tumor location (liver, lung, or subcutaneous). The decrease in γ-GCS activity was associated with lower intracellular GSH levels. Nrf2- and p53-dependent down-regulation of γ-GCS was associated with a decrease in the activities of superoxide dismutase 1 and 2, catalase, glutathione peroxidase, and glutathione reductase, but not of the O2--generating NADPH oxidase. The GCR knockdown-induced decrease in antioxidant protection caused a drastic decrease in the survival of metastatic cells during their interaction with endothelial cells, both in vitro and in vivo; only 10% of cancer cells attached to the endothelium survived compared to 90% survival observed in the controls. This very low rate of metastatic cell survival was partially increased (up to 52%) in vivo by inoculating B16-F10 cells preloaded with GSH ester, which enters the cell and delivers free GSH. Taken together, our results indicate that glucocorticoid signaling influences the survival of metastatic cells during their interaction with the vascular endothelium.
authors.affiliation: Department of Physiology, University of Valencia, Valencia, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Obrador
authors.affiliation: Department of Physiology, University of Valencia, Valencia, Spain.
authors.forename: Soraya L
authors.initials: SL
authors.lastname: Valles
authors.affiliation: Faculty of Experimental Sciences, San Vicente Martir Catholic University, Valencia, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Benlloch
authors.affiliation: Department of Physiology, University of Valencia, Valencia, Spain.
authors.forename: J Antoni
authors.initials: JA
authors.lastname: Sirerol
authors.affiliation: Department of Physiology, University of Valencia, Valencia, Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: Pellicer
authors.affiliation: Pathology Laboratory, Quirón Hospital, Valencia, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Alcácer
authors.affiliation: Pathology Laboratory, Quirón Hospital, Valencia, Spain.
authors.forename: Javier Alcácer-F
authors.initials: JA
authors.lastname: Coronado
authors.affiliation: Department of Physiology, University of Valencia, Valencia, Spain.
authors.forename: José M
authors.initials: JM
authors.lastname: Estrela
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D016923:Cell Death / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D015536:Down-Regulation / Q000235:genetics; D004703:Endocrine System / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D005721:Glutamate-Cysteine Ligase / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D008546:Melanoma, Experimental / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009362:Neoplasm Metastasis / Q000235:genetics*; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D011965:Receptors, Glucocorticoid; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione