title: [Enhanced reporter gene transfer and expression in cardiac myocytes mediated by ultrasonic destruction of the microbubbles].
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Cardiology, First Affiliated Hospital of Zhejiang University Medical School, Hangzhou 310031, China.
authors.forename: Guo-Zhong
authors.initials: GZ
authors.lastname: Wang
authors.forename: Shen-Jiang
authors.initials: SJ
authors.lastname: Hu
authors.forename: Zhe-Lan
authors.initials: ZL
authors.lastname: Zheng
authors.forename: Jian
authors.initials: J
authors.lastname: Sun
authors.forename: Jiang
authors.initials: J
authors.lastname: Li
authors.forename: Xia
authors.initials: X
authors.lastname: Zheng
authors.forename: Zhao-Hui
authors.initials: ZH
authors.lastname: Zhu
authors.forename: Yu-Mei
authors.initials: YM
authors.lastname: Yao
mesh_terms: D000818:Animals; D015870:Gene Expression; D017930:Genes, Reporter; D032383:Myocytes, Cardiac / Q000166:cytology*; D010957:Plasmids; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics