RESUMEN
This study investigated whether nicastrin can induce apoptotic cell death in SK-N-MC cells. MTT assays revealed the transfected cells expressing mutant nicastrin, compared with those expressing wild nicastrin or the control vector, showing significantly increased cell death. The mutant nicastrin transfectants were also observed to induce cytosolic cytochrome c release from the mitochondria, and Bax protein expression in response, to increased cell death. These observations suggested that nicastrin, as well as the APP and PS proteins, were also involved in the upregulated Bax mediated neuroblastoma cell death and the release of cytochrome c in the neuroblastoma.
Asunto(s)
Citocromos c/metabolismo , Regulación de la Expresión Génica , Glicoproteínas de Membrana/fisiología , Mutación , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Secretasas de la Proteína Precursora del Amiloide , Western Blotting/métodos , Muerte Celular/fisiología , Línea Celular Tumoral , Supervivencia Celular/fisiología , Análisis Mutacional de ADN/métodos , Humanos , Glicoproteínas de Membrana/genética , Mitocondrias/metabolismo , Mutagénesis Sitio-Dirigida/fisiología , Neuroblastoma , Reacción en Cadena de la Polimerasa/métodos , Sales de Tetrazolio , Tiazoles , Transfección/métodos , Proteína X Asociada a bcl-2RESUMEN
Estrogen influences the processing of the amyloid beta precursor protein (APP) in the pathogenesis of Alzheimer's disease, and this effect is mediated by estrogen receptors (ERs) in activating mitogen-activated protein kinase (MAPK)-signaling pathway. To test whether the estrogenic effect on both carboxyl-terminal amino acid fragment (C-terminal) of APP (APP-C105)- and ERbeta-mediated MAPK activation in in vitro, two hybrid genes containing each human ERbeta and APP-C105 gene fused to the neuron-specific enolase (NSE) promoter were constructed and were transfected to the neuronal SK-N-MC cells. Western blot shows that the activation of JNK-signaling pathway, but not p38 and ERK, is dependent on ERbeta through estrogen treatment and APP-C105 is also mediated through estrogen in activating MAPK-signaling pathway. The results suggest that ERbeta and APP-C105 derived from APP are necessary for estrogenic effect in activating MAPK-signaling pathway.