Microbubble-enhanced ultrasound exposure improves gene transfer in vascular endothelial cells

• Published in: World journal of gastroenterology : WJG 2006-12, Vol.12 (46), p.7508-7513
• Main Authors: Nie, Fang, Xu, Hui-Xiong, Tang, Qing, Lu, Ming-De
• Format: Article
• Language: English
• Subjects: Basic Research; Cells, Cultured; Dextrans; Endothelial Cells - metabolism; Fluorescein-5-isothiocyanate - analogs & derivatives; Green Fluorescent Proteins - genetics; Humans; Microbubbles; Phospholipids; Plasmids - genetics; Recombinant Proteins - genetics; Sulfur Hexafluoride; Transfection - methods; Ultrasonics; 细胞膜; 绿色荧光蛋白; 脐静脉上皮细胞; 质粒表达; 超声暴露; 转基因
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v12.i46.7508

AIM： To explore the effects of ultrasound exposure combined with microbubble contrast agent （SonoVue） on the permeability of the cellular membrane and on the expression of plasrnid DNA encoding enhanced green fluorescent protein （pEGFP） transfer into human umbilical vein endothelial cells （HUVECs）. METHODS： HUVECs with fluorescein isothiocyanatedextran （FD500） and HUVECs with pEGFP were exposed to continuous wave （1.9 MHz, 80.0 mW/cm^2） for 5 min, with or without a SonoVue. The percentage of FD500 taken by the HUVECs and the transient expression rate of pEGFP in the HUVECs were examined by fluorescence microscopy and flow cytornetry, respectively. RESULTS： The percentage of FDS00-positive HUVECs in the group of ultrasound exposure combined with SonoVue was significantly higher than that of the group of ultrasound exposure alone （24.0%± 5.5% vs 66.6% ± 4.1%, P 〈 0.001）. Compared with the group of ultrasound exposure alone, the transfection expression rate of pEGFP in HUVECs was markedly increased with the addition of SonoVue （16.1% ± 1.9% vs 1.5% ± 0.2%, P 〈 0.001）. No statistical significant difference was observed in the HUVECs survival rates between the ultrasound group with and without the addition of SonoVue （94.1% ± 2.3% vs 91.1% ± 4.1% ）. CONCLUSION： The cell membrane permeability of HUVECs and the transfection efficiency of pEGFP into HUVECs exposed to ultrasound are significantly increased after addition of an ultrasound contrast agent without obvious damage to the survival of HUVECs. This non- invasive gene transfer method may be a useful tool for clinical gene therapy of hepatic tumors.