Comparing transfection efficiency and safety for antisense oligodeoxyribonucleotide between phospholipids-based microbubbles and liposomes.
OBJECTIVE
To compare transfection efficiency and safety for antisense oligodeoxynucleotides (AS-ODNs) between two type of phospholipids-based vectors.


METHODS
An AS-ODNs sequence HA824 combined with luciferase reporter plasmid was used. Under low intensity ultrasound (US), a breast cancer cell line SK-BR-3 was exposed to different concentration of microbubbles and liposomes. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by propidium iodide assay. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the inhibitory effect of HA824 on HER-2 expression at mRNA level. Atomic force microscopy (AFM) scanning techniques was employed to observe the change of membrane pore size.


RESULTS
AS-ODNs transfection efficiency showed an increasing tend with microbubble concentration, but not with liposome concentration. Maximum transfection efficiency with minimum cell viability was achieved under 2% microbubble concentration. Too strong sonoporation activity would enlarge membrane pores significantly and cause low cell viability.


CONCLUSIONS
US-mediated AS-ODNs transfection enhanced by phospholipids-based microbubbles represents an effective and safe avenue.
Zhao
Ying-Zheng
YZ
Department of Clinical Pharmacology, General Hospital of Beijing Command of PLA, Beijing 100700, People's Republic of China. lctuua@yahoo.com.cn
Luo
Yu-Kun
YK
Liang
Hai-Dong
HD
Mei
Xing-Guo
XG
Tang
Jie
J
Lu
Cui-Tao
CT
Zhang
Yan
Y
Lin
Qian
Q
D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002470:Cell Survival / Q000187:drug effects; D004337:Drug Carriers; D005079:Excipients; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D018734:Genes, erbB-2 / Q000235:genetics; D006801:Humans; D008081:Liposomes; D008156:Luciferases / Q000235:genetics; D008863:Microspheres; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage / Q000009:adverse effects* / Q000494:pharmacology*; D010743:Phospholipids; D016062:Porosity; D011419:Propidium; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013054:Spectrophotometry, Atomic; D014162:Transfection
D004337:Drug Carriers; D005079:Excipients; D008081:Liposomes; D020319:Oligodeoxyribonucleotides, Antisense; D010743:Phospholipids; D012333:RNA, Messenger; D011419:Propidium; D008156:Luciferases