Effects ofultrasound-mediated SonoVue microbubbles destruction on the integrity and expression ofhAng-1 gene

Abstract

：Objective To test theefficiency of gene transfer and expression mediated by ultrasound and microbubble strategyin 293T cell. Methods SonoVue microbubbles were mixed with pla.smid DNA encoding hAng-1and green fluorescent protein. The mixture of the DNA and microbubbles was administer tocultured 293T cells by ultrasound exposue. The ultrasound condition was 1. 5 W/cm~2 and 30s. Forty eight hours later, transfer rate was assessed by fluorescence microscopy and flowcytometry. Cell viability was assayed by Trypan Blue staining. RT-PCR and Western blotanalysis was used to examine the expression of hAng-1 mRNA and protein. Agarose gelelectrophoresis was used to evaluate the integrity of the plasmid. Results Thetransfection expression rate of eGFP in 293T cells was markedly increased with the additonof 20% microbubbles and 15 mg/L DNA. Fetal calf serum had no influence on the genetransfer rate. Ultrasound irradiation combined with microbubbles couldn't destroy theintegrity of plasmid. Conclusions Ultrasound-mediated microbubbles destruction canincrease the transfection and expression of hAng-1 gene in 293T.