The improvement of liposome-mediated transfection of pEGFP DNA into             human prostate cancer cells by combining low-frequency and low-energy ultrasound             with microbubbles.
The aim of this study was to explore the use of a contrast agent to study             the effects of exposure to ultrasound, in combination with microbubbles, on liposome-mediated             transfection of genes into human prostate cancer cells. A contrast agent was used             to study the effects of ultrasound exposure in combination with microbubbles on             liposomes, which transfect genes into human prostate cancer cells. The human prostate             cancer cell line PC-3 in suspension was exposed to ultrasound with a 20% duty             cycle (i.e., 2 sec 'on' time and 8 sec 'off' time) lasting 5 min, with and without             ultrasound contrast agent (SonoVue™) using a digital sonifier at a frequency of             21 kHz and an intensity of 4.6 mW/cm2. Immediately after exposure to ultrasound,             cell viability and membrane damage were measured. After exposure to ultrasound,             the cell suspensions were put into 12‑well plates and cultured for 24 h. Fluorescence             microscopy and flow cytometry were used to detect pEGFP transfection efficiency.             Exposure to ultrasound alone and ultrasound combined with microbubbles resulted             in minimal cell death and induced negligible cell membrane damage. Ultrasound             combined with microbubbles had a greater effect on cell membrane damage in all             groups: the average cell membrane damage was 41.87%, and it was approximately             42‑fold greater than in the control group. The average transfection efficiency             of PC-3 cells was 20.30% for the liposome (Lipofectamine™)+pEGFP+ultrasound+ultrasound             contrast agent (SonoVue) group; this was the highest rate of all groups measured             and was approximately 81‑fold greater than that of the control group. The use             of low-frequency and low-energy ultrasound, in combination with microbubbles,             could be a potent physical method for increasing liposome gene delivery efficiency.             This technique is a promising non-viral approach that can be used in prostate             cancer gene therapy.
Department of Ultrasound in Medicine, Shanghai Jiao Tong University Affiliated 6th People's Hospital, Shanghai Institute of Ultrasound in Medicine, Shanghai 200233, PR China.
Wen-Kun
WK
Bai
Zuo-Hui
ZH
Wu
E
E
Shen
Ji-Zhen
JZ
Zhang
Bing
B
Hu
D016923:Cell Death; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000473:pathology / Q000648:ultrastructure; D017930:Genes, Reporter; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008081:Liposomes; D008297:Male; D045423:Microbubbles; D010743:Phospholipids; D010957:Plasmids; D011471:Prostatic Neoplasms / Q000628:therapy*; D013459:Sulfur Hexafluoride; D014162:Transfection / Q000379:methods*
D008081:Liposomes; D010743:Phospholipids; C420843:contrast agent BR1; D049452:Green Fluorescent Proteins; D013459:Sulfur Hexafluoride