RESUMEN

High doses of the organic nitrate glyceryl trinitrate (GTN), a nitric oxide (NO) donor, are known to trigger apoptosis in human cancer cells. Here, we show that such a cytotoxic effect can be obtained with subtoxic concentrations of GTN when combined with H89, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulphonamide.2HCl. This synergistic effect requires the generation of reactive oxygen species (ROS) from H89 and NO from GTN treatment that causes cGMP production and PKG activation. Furthermore, the GTN/H89 synergy was attenuated by inhibition of P2-purinergic receptors with suramin and competition with ATP/UDP. By down-regulating genes with antisense oligonucleotides, P2-purinergic receptors P2X3, P2Y1, and P2Y6 were found to have a role in creating this cytotoxic effect. Thus, H89 likely acts as an ATP mimetic synergizing with GTN to trigger apoptosis in aggressive cancer cells.

Asunto(s)

Neoplasias del Colon/patología , Isoquinolinas/química , Neoplasias/metabolismo , Nitroglicerina/química , Receptores Purinérgicos/metabolismo , Sulfonamidas/química , Adenosina Trifosfato/química , Animales , Apoptosis , Línea Celular Tumoral , Neoplasias del Colon/metabolismo , Sinergismo Farmacológico , Perfilación de la Expresión Génica , Humanos , Potencial de la Membrana Mitocondrial , Ratones , Óxido Nítrico/química , Oligonucleótidos Antisentido/química , Inhibidores de Proteínas Quinasas/química , Especies Reactivas de Oxígeno/química , Receptores Purinérgicos P2/metabolismo , Receptores Purinérgicos P2X3/metabolismo , Receptores Purinérgicos P2Y1/metabolismo , Transducción de Señal , Transfección