Enhanced reporter gene transfer and expression in cardiac myocytes mediated by ultrasonic destruction of the microbubbles / 中国应用生理学杂志
Chinese Journal of Applied Physiology
;
(6): 371-375, 2005.
Artículo
en Chino
| WPRIM
| ID: wpr-287006
ABSTRACT
<p><b>AIM</b>To determine if the diagnostic ultrasound and self-made microbubbles could be used to increase gene transfection and expression in cardiac myocytes by means of the ultrasound-mediated microbubbles destruction.</p><p><b>METHODS</b>The perfluoropropane-exposed sonicated dextrose albumin(PESDA) microbubbles were made and mixed with indicated volume reporter gene encoding beta-galactosidase prior to gene transfection. Gene transfection into the cultured cardiac myocytes was performed by exposure to the various intense diagnostic ultrasound (1.3 MHz) in the presence of the gene-attached microbubbles. The calcium phosphate precipitation gene transfection was carried out alone or in combination with ultrasound-mediated destruction microbubbles. The cells were harvested 48 h after transfection and beta-galactosidase expression was detected by in situ staining and quantitive assay.</p><p><b>RESULTS</b>Cardiac myocytes exposed to ultrasound with PESDA induced significantly increase in gene expression (60-fold compared with naked plasmids transfection, P < 0.01). Moreover, it was found that the reporter gene expression not only related with ultrasound intension but also with the microbubbles concentration. In combination with calcium phosphate precipitation gene transfection, ultrasound-mediated destruction microbubbles resulted in more intense gene expression even 6 hours after calcium phosphate precipitation gene transfection.</p><p><b>CONCLUSION</b>The ultrasonic destruction of gene-loaded microbubble is a highly effective gene transfer method, and it not only acts on the gene entry into cells, but also on the intracellular exogenous DNA expression.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Plásmidos
/
Ultrasonido
/
Transfección
/
Expresión Génica
/
Ratas Wistar
/
Genes Reporteros
/
Biología Celular
/
Miocitos Cardíacos
/
Métodos
Límite:
Animales
Idioma:
Chino
Revista:
Chinese Journal of Applied Physiology
Año:
2005
Tipo del documento:
Artículo
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