Application of ultrasonic gas-filled liposomes in enhancing transfer for breast cancer-related antisense oligonucleotides: an experimental study.
The aim of this study was to investigate the application of ultrasonic gas-filled liposomes in enhancing transfer for breast cancer-related antisense oligonucleotides in vitro. An antisense oligodeoxynucleotide (AS-ODN) sequence, HA2741, modified with luciferase reporter plasmid, was used in evaluating the enhancing effect of gas-filled liposomes for gene transfer in breast cancer cells. Some important factors on HA 2741 transfection efficiency, such as wave intensity, ultrasound duration, gas-filled liposome concentration, and HA2741 concentration, were tested, respectively. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by propidium iodide assay. Reverse-transcriptase polymerase chain reaction and immunocytochemistry were used to detect the inhibitory effect of HA2741 on HER-2 expression. All the four factors (wave intensity, ultrasound duration, gas-filled liposome concentration, and HA2741 concentration) showed a positive effect on AS-ODN transfection efficiency. However, these factors had a negative effect on cell viability. Considering all the factors investigated, the maximum transfection efficiency with minimum cell viability achieved under 2% gas-filled liposome mixed with 80 nmol/L HA2741 for 30-second ultrasound exposure at -3.0 dB wave intensity, which gave an overall transfection efficiency exceeding 90% and a cell viability near 90%. Under controlled conditions, ultrasound-mediated AS-ODN transfer, enhanced by gas-filled liposomes, may represent an effective, safe avenue for cancer-related gene delivery.
Luo
Yu-Kun
YK
Department of Ultrasound, General Hospital of PLA, Beijing, China.
Zhao
Ying-Zheng
YZ
Lu
Cui-Tao
CT
Tang
Jie
J
Li
Xiao-Kun
XK
D001943:Breast Neoplasms / Q000235:genetics / Q000628:therapy; D045744:Cell Line, Tumor; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D045423:Microbubbles; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D014465:Ultrasonics
D008081:Liposomes; D016376:Oligonucleotides, Antisense; D018719:Erb-b2 Receptor Tyrosine Kinases