Aim To develop a novel method to effectively deliver enhanced green fluorescence protein (EGFP)C3 into Chinese hamster orary (CHO) cells in vitro by ultrasound-mediated microbubble destruction. Methods Expression of the EGFPC3 gene was quantified by laser confocal microscopy and FACS analysis. Cell viability was assayed by Trypan Blue staining. Results Ultrasound combined with microbubbles can enhance gene transfer in cultured cells, but the effect is vary according to the utrasound condition and concetration of microbubble. Optimal gene expression occurred with microbubble at the concentration of 10% and ultrasound parameter at 0.8 MHz, 1.0 W/cm 2,10% duty cycle, 60 s. Under this condition, the transfection rate of EGFPC3 gene with ultrasound-mediated microbubble destruction method was similar to that of transfection with lipofectamine (56.2%±2.6% versus 60.8%±4.1%, p>0.05) and the relative fluorescence intensity of EGFPC3 gene was as high as that of transfection with lipofectamine (2035±32 versus 2140±28, p>0.05). Furthermore both albumin microbubble and ultrasound had no effect on cell viability. The cell viablity were 94.1%±4.6% or 93.8%±3.1% when cultured with 10% albumin microbubble or under ultrasound at 0.8 MHz, 1.0 W/cm 2, 10% duty cycle, 60 s respectively, which were no significantly difference compared with controls. Conclusions These results suggest a possible new strategy for ultrasound-mediated microbubble destruction method in gene therapy.