Cisplatin cytotoxicity in organ of Corti-derived immortalized cells.
Cisplatin is an anticancer drug currently used in the treatment of genital and head and neck tumors. Its use in these and other types of tumors is narrowed by onset of chemoresistance and severe undesired side effects, like as nephro- and ototoxicity, whose mechanisms of action are only partially understood. In the present study we investigated the effects of cisplatin (cis-dichlorodiaminoplatin, CDDP) on a cell line (OC-k3) developed from organs of Corti of transgenic mice. We observed at 48 h that cell death due to cisplatin was time and concentration-dependent. The cell death displayed some morphological hallmarks of apoptosis, including nuclear fragmentation into several large nuclear fragments, surrounded by a rearranged and thickened actin cytoskeleton. No DNA laddering was detected, suggesting absence of endonuclease activity, nor annexin V positivity, suggesting absence of phosphatidylserine externalization. Several molecules protected the cells against CDDP induced cytotoxicity, including methionine, suramin and PD98059. Methionine reduced CDDP-uptake, while suramin, a polycathionic compound a specifically binding external proteins, did not. This finding suggested that suramin could exert its protective effect by acting on an intracellular transduction pathway. We tested this hypothesis by studying the effect of suramin and PD98059, a MEK inhibitor, on the mitogen activated protein kinase (MAPK) cascade. After CDDP treatment, we found an increase of phosphorylation of extracellular regulated kinases (ERK)1/2, that could be inhibited by PD98059 and suramin. These data suggest that ERK pathways can play a role in mediating the cell death induction in presence of a CDDP challenge.
Previati
Maurizio
M
Department of Morphology and Embryology, Human Anatomy Division, University of Ferrara, Ferrara, Italy. prm@unife.it
Lanzoni
Irene
I
Astolfi
Laura
L
Fagioli
Francesco
F
Vecchiati
Giorgio
G
Pagnoni
Antonella
A
Martini
Alessandro
A
Capitani
Silvano
S
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002461:Cell Line, Transformed; D002945:Cisplatin / Q000633:toxicity*; D019610:Cytoprotection / Q000187:drug effects; D004247:DNA / Q000302:isolation & purification; D004789:Enzyme Activation / Q000187:drug effects; D008715:Methionine / Q000494:pharmacology; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D009925:Organ of Corti / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D010766:Phosphorylation / Q000187:drug effects; D011419:Propidium / Q000378:metabolism; D013498:Suramin / Q000494:pharmacology
D000970:Antineoplastic Agents; D011419:Propidium; D013498:Suramin; D004247:DNA; D008715:Methionine; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D053148:Caspase 3; D002945:Cisplatin