Objective To evaluate inhibitory effect of retinoblastoma（RB） by ultrasound targeted carry HSV1-tk targeted microbubble destruction. Methods Carring HSV1-tk and VEGFR2 antibody targeted microbubble was Prepared. All the RB cells were divided into 7 groups：control group，cells+plasmid，cells+plasmid+SonoVue，cells+targeted microbubble，cells+plasmid+ultrasound，cells+plasmid+SonoVue+ultrasound，cells+targeted microbubble+ultrasound. Gene transfection results were observed under inversion fluorescent microscope. Seventy–two hours later，gene transfection rates were guantified under flow cytometry. Inhibitory rate of RB cells were evaluated after GCV was added. Results The transfection rate of cells+targeted microbubble+ultrasound group was（24.78±1.04）%，which was higher than cells+plasmid+SonoVue+ultrasound group（14.31±0.69）%. With the increase of GCV concentration and the prolongation of culture time, the inhibition rate of each group increased gradually. When the GCV concentration was 100mg/l，afer ninety–six hours culture time， inhibitory rate of cells+targeted microbubble+ultrasound group were （92.91±1.71）%. Conclusion Ultrasound targeted carry HSV1-tk targeted microbubble destruction can effectively inhibit RB cells，after GCV was added.