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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.
Article in Chinese | 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>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Plasmids / Ultrasonics / Transfection / Gene Expression / Rats, Wistar / Genes, Reporter / Cell Biology / Myocytes, Cardiac / Methods Limits: Animals Language: Chinese Journal: Chinese Journal of Applied Physiology Year: 2005 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Plasmids / Ultrasonics / Transfection / Gene Expression / Rats, Wistar / Genes, Reporter / Cell Biology / Myocytes, Cardiac / Methods Limits: Animals Language: Chinese Journal: Chinese Journal of Applied Physiology Year: 2005 Type: Article