[Echogenic phospholipids-based gas-filled microbubbles as delivery system of antisense oligodeoxynucleotides].
OBJECTIVE
To investigate the feasibility of transfer antisense oligodeoxynucleotides (AS-ODNs) by the phospholipids-based gas-filled microbubbles (PGM) under ultrasound activation.


METHODS
An antisense oligodeoxynucleotides sequence ZL combined with luciferase reporter plasmid was used. A breast cancer cell line SK-BR-3 was exposed to different conditions to investigate the effects of such factors as ZL concentration, PGM concentration, mechanical index (MI) and ultrasound exposure duration on transfection efficiency and cell viability. The transfection efficiency and cell viability by other lipid vectors such as lipofectamine and liposome were also tested, whose results were comparied with that of PGM. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by PI (propidium iodide) assay.


RESULTS
Among the factors tested, ultrasound exposure duration, MI and PGM concentration had obvious impacts on transfection efficiency and cell viability. The results showed that the optimal ultrasound condition was the exposure to ultrasound at MI 1.0 for 30 s with 2% PGM concentration, which gave an overall transfection efficiency of 78% +/- 10%, increased nearly 18 folds over the transfection by PGM (4.0%) or lipofectamine (4.3%) without ultrasound. Under same ultrasound conditions, different vectors showed significant difference in transfection efficiency while there are similar results in cell viability.


CONCLUSIONS
Under proper ultrasound conditions, PGM can markedly enhance AS-ODNs transfection efficiency.
Zhao
Ying-zheng
YZ
Department of Clinical Pharmacology, General Hospital of Beijing Command, PLA, Beijing 100700, China.
Luo
Yu-kun
YK
Tang
Jie
J
Mei
Xing-guo
XG
Zhang
Yan
Y
Lin
Qian
Q
D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival; D004337:Drug Carriers; D005260:Female; D006801:Humans; D008055:Lipids; D008081:Liposomes; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D045423:Microbubbles; D008856:Microscopy, Fluorescence; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics*; D010743:Phospholipids; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics
D004337:Drug Carriers; D008055:Lipids; C086724:Lipofectamine; D008081:Liposomes; D020319:Oligodeoxyribonucleotides, Antisense; D010743:Phospholipids; D008156:Luciferases