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Effect of silencing myocardin gene expression on differentiation of mouse bone mesenchymal stem cells into smooth muscle-like cells induced by PDGF-BB / 中华病理学杂志
Chinese Journal of Pathology ; (12): 117-120, 2009.
Artículo en Chino | WPRIM | ID: wpr-319775
ABSTRACT
<p><b>OBJECTIVE</b>Construction of a small interfering RNA (siRNA) eukaryotic expression vector specific to mouse myocardin gene and study on the role of myocardin-siRNA on differentiation of mouse bone mesenchymal stem cells (MSCs) into smooth muscle-like cells induced by PDGF-BB in vitro.</p><p><b>METHODS</b>Mouse MSCs were isolated from bone marrow and cultured with 50 mg/L PDGF-BB and fetal bovine serum (20%). Specific myocardin-siRNA sequence was cloned into a plasmid pGenesil-1.0 vector, which contained U6 promoter. The recombinant plasmid and control plasmid were transfected into MSCs which had been cultured with PDGF-BB for 6 days beforehand. The expression of myocardin mRNA was detected by RT-PCR 48 hours after the transfection. Immunohistochemistry was used to detect the SM-MHC and to identify the smooth muscle-like cells.</p><p><b>RESULTS</b>The recombinant plasmids carrying myocardin-siRNA sequences were constructed successfully and the myocardin mRNA was reduced 42.86% by pGen-myo-shRNA in comparing with that of the controls (P<0.01); and the expression of SM-MHC protein was down-regulated (P<0.01).</p><p><b>CONCLUSION</b>Subset of mouse MSCs have the potential to differentiate into smooth muscle-like cells, a possible cell source responsible for atherosclerotic plaque formation, and myocardin expression may play an important role during this process.</p>
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Patología / Farmacología / Fisiología / Plásmidos / Factor de Crecimiento Derivado de Plaquetas / ARN Mensajero / Células de la Médula Ósea / Proteínas Nucleares / Transfección / Regulación hacia Abajo Tipo de estudio: Estudio pronóstico Límite: Animales Idioma: Chino Revista: Chinese Journal of Pathology Año: 2009 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Patología / Farmacología / Fisiología / Plásmidos / Factor de Crecimiento Derivado de Plaquetas / ARN Mensajero / Células de la Médula Ósea / Proteínas Nucleares / Transfección / Regulación hacia Abajo Tipo de estudio: Estudio pronóstico Límite: Animales Idioma: Chino Revista: Chinese Journal of Pathology Año: 2009 Tipo del documento: Artículo