title: Cardiac glycoside-induced cell death and Rho/Rho kinase pathway: Implication of different regulation in cancer cell lines.
abstract: Previously, we demonstrated that the Rho/ROCK pathway is involved in ouabain-induced apoptosis in HUVEC. In the current work, we investigated whether the Rho/ROCK pathway is functional during cardiac glycosides-induced cytotoxic effects in cancer cell lines, as well as in non-tumor cells. For that purpose, we evaluated the role of ROCK activation in bleb formation and cell migration over upstream and downstream effectors in addition to ROCK cleavage after cardiac glycosides treatment. All three cardiac glycosides (ouabain, digoxin and bufalin) induced cell death in HeLa and HepG2 cells and increased the formation of blebbing in HeLa cells. In contrast to our previous study, ROCK inhibitor Y27632 did not prevent bleb formation. Observation of ROCK II cleavage after ouabain, digoxin and oxaliplatin treatments in HeLa and/or HepG2 cells suggested that cleavage is independent of cell type and cell death induction. While inhibiting cleavage of ROCK II by the caspase inhibitors z-VAD-fmk, z-VDVAD-fmk and z-DEVD-fmk, evaluation of caspase 2 siRNA ineffectiveness on this truncation indicated that caspase-dependent ROCK II cleavage is differentially regulated in cancer cell lines. In HeLa cells, ouabain induced the activation of ROCK, although it did not induce phosphorylation of ERM, an upstream effector. While Y27632 inhibited the migration of HeLa cells, 10nM ouabain had no effect on cell migration. In conclusion, these findings indicate that the Rho/ROCK pathway is regulated differently in cancer cell lines compared to normal cells during cardiac glycosides-induced cell death.
authors.affiliation: Department of Pharmacology, Gazi University, Faculty of Pharmacy, 06330, Turkey.
authors.forename: Aysun
authors.initials: A
authors.lastname: Özdemir
authors.affiliation: Department of Pharmacology, Gazi University, Faculty of Pharmacy, 06330, Turkey.
authors.forename: Yaprak Dilber
authors.initials: YD
authors.lastname: Şimay
authors.affiliation: Department of Pharmacology, Gazi University, Faculty of Pharmacy, 06330, Turkey.
authors.forename: Burçin
authors.initials: B
authors.lastname: İbişoğlu
authors.affiliation: Department of Pharmacology, Gazi University, Faculty of Pharmacy, 06330, Turkey.
authors.forename: Biljana
authors.initials: B
authors.lastname: Yaren
authors.affiliation: Department of Pharmacology, Gazi University, Faculty of Pharmacy, 06330, Turkey.
authors.forename: Döne
authors.initials: D
authors.lastname: Bülbül
authors.affiliation: Department of Pharmacology, Gazi University, Faculty of Pharmacy, 06330, Turkey. Electronic address: mark@gazi.edu.tr.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Ark
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002301:Cardiac Glycosides / Q000494:pharmacology*; D053143:Caspase 2 / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects / Q000473:pathology; D004789:Enzyme Activation / Q000187:drug effects; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D009336:Necrosis / Q000139:chemically induced; D010042:Ouabain / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D059748:Proteolysis / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects*; D020741:rho GTP-Binding Proteins / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: D002301:Cardiac Glycosides; D010042:Ouabain; D054460:rho-Associated Kinases; D053143:Caspase 2; D020741:rho GTP-Binding Proteins
keywords: Cancer; Cardiac glycosides; Cell death; Migration; Rho kinase