title: Ultrasound-mediated microbubble destruction enhances gene transfection in pancreatic cancer cells.
abstract: BACKGROUND
  The purpose of this study was to determine whether ultrasound exposure combined with microbubble destruction could be used to enhance non-viral gene delivery in human pancreatic carcinoma cells (PANC-1).
  
  
  METHODS
  The study was performed with four experimental groups: Group P, plasmid alone; Group P+M, plasmid and microbubbles; Group P+U, plasmid and ultrasound; Group P+U+M, plasmid with ultrasound and microbubbles. Plasmid DNA encoding enhanced green fluorescent protein (pEGFP) was gently mixed with commercially available ultrasound microbubble contrast agents (SonoVue; Bracco Diagnostics Inc, Milan, Italy) in Group P+M and Group P+U+M. The different combinations of DNA and DNA plus microbubbles were added to cultured PANC-1 cells under different conditions. Transfection efficiency and cell viability were assessed by FACS analysis (Becton Dickinson, San Jose, CA, USA), confocal laser scanning microscopy, and trypan blue staining.
  
  
  RESULTS
  The results demonstrated that microbubbles with ultrasound exposure could significantly enhance the reporter gene expression as compared with other groups (Group P+U+M, 21.4%+/-3.16%; Group P, 2.9%+/-0.45%; Group P+M, 3.1%+/-0.51%; Group P+U, 6.1%+/-1.27%; P<0.01). No statistically significant difference was observed in the PANC-1 cell viability between Group P+U+M and other groups (P>0.05).
  
  
  CONCLUSIONS
  Our in-vitro findings suggest that ultrasound-mediated microbubble destruction has the potential to promote efficient gene transfer into PANC-1 cells without significant cell death. This non-invasive gene transfer method may be a useful tool for safe clinical gene therapy of pancreatic cancer in the future.
authors.affiliation: Department of Ultrasound, First Affiliated Hospital of Harbin Medical University, Harbin, China. wangjunfeng2008@gmail.com
authors.forename: Jun-Feng
authors.initials: JF
authors.lastname: Wang
authors.forename: Jia-Bei
authors.initials: JB
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Chen
authors.forename: Chun-Mei
authors.initials: CM
authors.lastname: Zhang
authors.forename: Lan
authors.initials: L
authors.lastname: Liu
authors.forename: Shang-Ha
authors.initials: SH
authors.lastname: Pan
authors.forename: Chang-Jun
authors.initials: CJ
authors.lastname: Wu
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival; D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D045423:Microbubbles; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000628:therapy; D010743:Phospholipids; D010957:Plasmids / Q000235:genetics; D013459:Sulfur Hexafluoride; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D014465:Ultrasonics
chemical_list: D010743:Phospholipids; C420843:contrast agent BR1; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013459:Sulfur Hexafluoride