Effects of ultrasound-mediated SonoVue microbubbles destruction on the integrity and expression of hAng-1 gene
10.3760/cma.j.issn.1004-4477.2009.12.022
- VernacularTitle:超声介导SonoVue微泡破裂对hAng-1基因完整性和表达效率的影响
- Author:
Qian CHEN
;
Qing ZHOU
;
Yinghui WANG
;
Xiao WANG
;
Ruiqiang GUO
- Publication Type:Journal Article
- Keywords:
Ultrasonography;
Microbubble;
Transfection;
Myocardial ischemia
- From:
Chinese Journal of Ultrasonography
2009;18(12):1080-1084
- CountryChina
- Language:Chinese
-
Abstract:
Objective To test the efficiency of gene transfer and expression mediated by ultrasound and microbubble strategy in 293T cell. Methods SonoVue microbubbles were mixed with pla.smid DNA encoding hAng-1 and green fluorescent protein. The mixture of the DNA and microbubbles was administer to cultured 293T cells by ultrasound exposue. The ultrasound condition was 1. 5 W/cm~2 and 30 s. Forty eight hours later, transfer rate was assessed by fluorescence microscopy and flow cytometry. Cell viability was assayed by Trypan Blue staining. RT-PCR and Western blot analysis was used to examine the expression of hAng-1 mRNA and protein. Agarose gel electrophoresis was used to evaluate the integrity of the plasmid. Results The transfection expression rate of eGFP in 293T cells was markedly increased with the additon of 20% microbubbles and 15 mg/L DNA. Fetal calf serum had no influence on the gene transfer rate. Ultrasound irradiation combined with microbubbles couldn't destroy the integrity of plasmid. Conclusions Ultrasound-mediated microbubbles destruction can increase the transfection and expression of hAng-1 gene in 293T.
