Variantes genéticas en el locus 9p21 contribuyen al desarrollo de arteriosclerosis a través de la modulación de ANRIL y CDKN2A/B

Registro creado en correspondencia al grado de doctora de Ada Congrains Castillo.Los estudios de asociación de todo el genoma (GWAS) han identificado variantes genéticas que contribuyen al riesgo de enfermedad cardiovascular (ECV) en el locus del cromosoma 9p21. La región asociada a CVD es adyacente a los dos inhibidores de quinasas dependientes de ciclina (CDKN) 2A y 2B y los últimos exones del ARN no codificante, ANRIL. Todavía no está claro cuál de estas transcripciones o cómo están involucradas en la patogénesis de la aterosclerosis.Genome-wide association studies (GWAS) have identified genetic variants contributing to the risk of cardiovascular disease (CVD) at the chromosome 9p21 locus. The CVD-associated region is adjacent to the two cyclin dependent kinase inhibitors (CDKN)2A and 2B and the last exons of the non-coding RNA, ANRIL. It is still not clear which of or how these transcripts are involved in the pathogenesis of atherosclerosis.Japón. Programa de Promoción de Estudios Fundamentales en el Instituto Nacional de Innovación Biomédica de Japón (HR: 22-2-5), el Ministerio de Educación, Cultura, Deportes, Ciencia y Tecnología de Japón (KK: 22510211) y la Fundación NOVARTIS para la Investigación Gerontológica (KK).Tesi

5α-Oleandrin reduce Bcl-2 protein and increase Bax protein expression on Hela cervical cancer cell

BACKGROUND The leaves of Nerium indicum Mill. have been utilized traditionally to cure cancer. By using Bioassay guided extraction and isolation method, three compounds (NiO-1, NiO-2, NiO-3) were isolated from an active fraction of the CHCl3 extract of N. indicum leaves. NiO-2 identified as 5α- oleandrin is the best cytotoxic compound on HeLa cervical cancer cell in vitro among the other two. However mechanism of action of the compound hasn’t been evaluated yet. The aims of this study were to determine the mechanism of action at molecular level. METHODS The action mechanism of 5α-oleandrin on HeLa cervical cancer cells was analyzed by staining the cells with Hoechst 33342; the agarose gel electrophoresis was aimed to determine the DNA fragmentation and the western blotting was aimed to determine the Bcl-2 and Bax protein expression. RESULTS Incubation of HeLa cervical cancer cell with 5α-oleandrin at the concentration 3,47x10-4 mM (24 hours) followed by staining with Hoechst 33342, a broken up light blue color of nucleus was observed (compared with intensive color of untreated control). By gel electrophoresis (at the same concentration of the tested compound), a smear band at about 200 bp was observed. In addition, cells treated with 5α-oleandrin displayed a decreasing of the Bcl-2 protein expression and increasing of the Bax protein expression CONCLUSION 5α-oleandrin induced HeLa cervical cancer cells DNA fragmentation observed by the presence of a smear band at about 200 bp indicative the apoptotic occurrence. 5α-oleandrin induces apoptosis by reducing the Bcl-2 protein expression but the Bax protein expression increases