Ultrasound-mediated microbubble destruction enhances gene transfection in pancreatic cancer cells.
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.
Wang
Jun-Feng
JF
Department of Ultrasound, First Affiliated Hospital of Harbin Medical University, Harbin, China. wangjunfeng2008@gmail.com
Wang
Jia-Bei
JB
Chen
Hua
H
Zhang
Chun-Mei
CM
Liu
Lan
L
Pan
Shang-Ha
SH
Wu
Chang-Jun
CJ
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
D010743:Phospholipids; C420843:contrast agent BR1; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013459:Sulfur Hexafluoride