title: Activation of dual apoptotic pathways in human melanocytes and protection by survivin.
abstract: Apoptosis resistance in melanoma is a primary cause of treatment failure. Apoptotic pathways in melanocytes, from which melanoma arises, are poorly characterized. Human melanocytes were susceptible to apoptosis following exposure to UV radiation (UVB, 24-48 hours), 4-tert-butylphenol (4-TBP, 1-4 hours), and cisplatin (24-48 hours). These responses were associated with Bid cleavage, caspase activation (caspases 3, 8, and 9), mitochondrial depolarization and release of cytochrome c, Smac/DIABLO, and apoptosis-inducing factor (AIF), but not endonuclease G. The apoptotic responses and AIF release were caspase-independent, as they were not blocked by zVal-Ala-Asp(OMe)-fluoromethyl ketone (zVAD-fmk). While RNA interference-mediated knockdown of AIF protected melanocytes against apoptosis induced by serum withdrawal, apoptotic responses to UVB, cisplatin, and 4-TBP were not compromised by AIF knockdown, even in the presence of zVAD-fmk. Finally, adenoviral-mediated expression of Survivin, an inhibitor of apoptosis expressed in melanoma but not melanocytes, protected melanocytes against UVB-induced apoptosis. Survivin expression in melanocytes partially blocked caspase activation and release of mitochondrial release of AIF, cytochrome c, and Smac induced by UVB. These data indicate that multiple stimuli can activate both caspase-dependent and caspase-independent apoptotic pathways in melanocytes, and that endogenous expression of Survivin in melanoma may contribute to apoptosis resistance by multiple mechanisms.
authors.affiliation: Huntsman Cancer Institute, University of Utah Health Sciences Center, Salt Lake City, 84112, USA.
authors.forename: Tong
authors.initials: T
authors.lastname: Liu
authors.forename: Diana
authors.initials: D
authors.lastname: Biddle
authors.forename: Adrianne N
authors.initials: AN
authors.lastname: Hanks
authors.forename: Brook
authors.initials: B
authors.lastname: Brouha
authors.forename: Hui
authors.initials: H
authors.lastname: Yan
authors.forename: Ray M
authors.initials: RM
authors.lastname: Lee
authors.forename: Sancy A
authors.initials: SA
authors.lastname: Leachman
authors.forename: Douglas
authors.initials: D
authors.lastname: Grossman
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis; D051033:Apoptosis Inducing Factor / Q000502:physiology; D053148:Caspase 3; D020169:Caspases / Q000502:physiology; D019610:Cytoprotection; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008544:Melanocytes / Q000166:cytology*; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D009363:Neoplasm Proteins / Q000502:physiology*; D000077022:Survivin
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D051033:Apoptosis Inducing Factor; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D000077022:Survivin; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases