title: The cytotoxic effects of VE-3N, a novel 1,4-dihydropyridine derivative, involve the mitochondrial bioenergetic disruption via uncoupling mechanisms.
abstract: Several 1,4-dihydropyridine derivatives overcome the multidrug resistance in tumors, but their intrinsic cytotoxic mechanisms remain unclear. Here we addressed if mitochondria are involved in the cytotoxicity of the novel 1,4-dihydropyridine derivative VE-3N [ethyl 6-chloro-5-formyl-2-methyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3-carboxylate] towards cancer cells by employing hepatic carcinoma (HepG2) cells and isolated rat liver mitochondria. In HepG2 cells, VE-3N induced mitochondrial membrane potential dissipation, ATP depletion, annexin V/propidium iodide double labeling, and Hoechst staining; events indicating apoptosis induction. In isolated rat liver mitochondria, VE-3N promoted mitochondrial uncoupling by exerting protonophoric actions and by increasing membrane fluidity. Mitochondrial uncoupling was evidenced by an increase in resting respiration, dissipation of mitochondrial membrane potential, inhibition of Ca2+uptake, stimulation of Ca2+release, decrease in ATP synthesis, and swelling of valinomycin-treated organelles in hyposmotic potassium acetate media. Furthermore, uncoupling concentrations of VE-3N in the presence of Ca2+plus ruthenium red induced the mitochondrial permeability transition process. These results indicate that mitochondrial uncoupling is potentially involved in the VE-3N cytotoxic actions towards HepG2 cells. Considering that hepatocellular carcinoma is the most common form of liver cancer, our findings may open a new avenue for the development of VE-3N-based cancer therapies, and help to unravel the cytotoxic mechanisms of 1,4-dihydropyridines towards cancer cells.
authors.affiliation: Center for Research and Biological Evaluations, Institute of Pharmacy and Food, University of Havana, 222 St. # 2317, La Coronela, La Lisa, Havana, Cuba.
authors.forename: Javier
authors.initials: J
authors.lastname: Marín-Prida
authors.affiliation: Center for Research and Biological Evaluations, Institute of Pharmacy and Food, University of Havana, 222 St. # 2317, La Coronela, La Lisa, Havana, Cuba. Electronic address: gpardo@ifal.uh.cu.
authors.forename: Gilberto L
authors.initials: GL
authors.lastname: Pardo Andreu
authors.affiliation: Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. Café s/n, 14040-903, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Camila Pederiva
authors.initials: CP
authors.lastname: Rossignoli
authors.affiliation: Center for Research and Biological Evaluations, Institute of Pharmacy and Food, University of Havana, 222 St. # 2317, La Coronela, La Lisa, Havana, Cuba.
authors.forename: Michael González
authors.initials: MG
authors.lastname: Durruthy
authors.affiliation: Laboratory of Organic Synthesis, Faculty of Chemistry, University of Havana, Zapata st./G and Carlitos Aguirre, Vedado Plaza de la Revolución, PO 10400, Havana, Cuba.
authors.forename: Estael Ochoa
authors.initials: EO
authors.lastname: Rodríguez
authors.affiliation: Laboratory of Organic Synthesis, Faculty of Chemistry, University of Havana, Zapata st./G and Carlitos Aguirre, Vedado Plaza de la Revolución, PO 10400, Havana, Cuba.
authors.forename: Yamila Verdecia
authors.initials: YV
authors.lastname: Reyes
authors.affiliation: Department of Pharmacy, Institute of Pharmacy and Food, University of Havana, 222 St. # 2317, La Coronela, La Lisa, Havana, Cuba.
authors.forename: Roberto Fernández
authors.initials: RF
authors.lastname: Acosta
authors.affiliation: Department of Clinical Analyses, Toxicology and Food Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. Café s/n, 14040-903 Ribeirão Preto, São Paulo, Brazil.
authors.forename: Sergio A
authors.initials: SA
authors.lastname: Uyemura
authors.affiliation: Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. Café s/n, 14040-903, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Luciane C
authors.initials: LC
authors.lastname: Alberici
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D004095:Dihydropyridines / Q000494:pharmacology*; D004734:Energy Metabolism / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014475:Uncoupling Agents / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D004095:Dihydropyridines; D014475:Uncoupling Agents; C000626605:ethyl 6-chloro-5-formyl-2-methyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3-carboxylate; D000255:Adenosine Triphosphate
keywords: Cancer; Cytotoxicity; Dihydropyridine; HepG2 cells; Mitochondria; Uncoupling