title: Staurosporine-induced apoptosis in Chang liver cells is associated with down-regulation of Bcl-2 and Bcl-XL.
abstract: A potent inhibitor of serine/threonine kinases, staurosporine exerts antiproliferative and apoptotic effects in many cancer cells, although the exact mechanism of its action is still unclear. This study examines the effects of staurosporine on Chang liver cells, an immortalized non-tumor cell line, in comparison with those caused in HuH-6 and HepG2 cells, two human hepatoma cell lines. Our results provide evidence that staurosporine promotes apoptosis in Chang liver cells as observed by flow cytometric analysis and acridine orange/ethidium bromide staining. The effect appeared already after 8 h of treatment and increased with treatment time and dose. After 48 h of exposure to 200 nM staurosporine clear apoptotic signs were observed in about 50% of the cells. Western blotting analysis showed that in Chang liver cells staurosporine induced a marked decrease in the levels of the antiapoptotic factors Bcl-2 (-75%) and Bcl-XL (-50%). Staurosporine also caused loss of mitochondrial transmembrane potential, release of cytochrome c from mitochondria and activation of caspase-3. The involvement of caspases in staurosporine-induced cell death was also suggested by the observation that the addition of z-VAD-fmk, a general inhibitor of caspases, suppressed apoptosis. In HuH-6 and HepG2 cells treatment with staurosporine induced the arrest of cells in G2/M phase of cell cycle. This effect was not modified by z-VAD-fmk and was not accompanied by the appearance of biochemical signs of apoptosis. We conclude that staurosporine induced apoptosis in Chang liver cells by a mitochondria-caspase-dependent pathway which was closely correlated with a decrease in Bcl-2 and Bcl-XL levels, while in HuH-6 and HepG2 hepatoma cells the drug caused only an antiproliferative effect.
authors.affiliation: Dipartimento di Biologia Cellulare e dello Sviluppo, Sezione di Biochimica, Università di Palermo, Policlinico, Palermo, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Giuliano
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Bellavia
authors.forename: Marianna
authors.initials: M
authors.lastname: Lauricella
authors.forename: Antonella
authors.initials: A
authors.lastname: D'Anneo
authors.forename: Barbara
authors.initials: B
authors.lastname: Vassallo
authors.forename: Renza
authors.initials: R
authors.lastname: Vento
authors.forename: Giovanni
authors.initials: G
authors.lastname: Tesoriere
mesh_terms: D000165:Acridine Orange / Q000494:pharmacology; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004996:Ethidium / Q000494:pharmacology; D005434:Flow Cytometry; D016195:G2 Phase; D006801:Humans; D008099:Liver / Q000166:cytology*; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D008938:Mitosis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis*; D019311:Staurosporine / Q000494:pharmacology*; D013997:Time Factors; D051020:bcl-X Protein
chemical_list: D000590:Amino Acid Chloromethyl Ketones; C494811:BCL2L1 protein, human; D004791:Enzyme Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D051020:bcl-X Protein; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D004996:Ethidium; D000165:Acridine Orange; D019311:Staurosporine