Experimental Study Of Wild-Type P53 Gene Transfected Into The Retinoblastoma Of Nude Mice By Ultrasound Microbuble Intensifier

Background and Objective:Retinoblastoma is the commonest intraocular maligant tumor in children.Nowadays,the therapy methods include picking off eyeball,laser photocoagulation,transpupillary thermotherapy,cryotherapy,ratiotherapy et al.Gene therapy for RB is still at the starting stage.As a new gene vetor,microbubble can break up and release the plasmid or drugs it carries at the appointed place by ultrasound.In the study,we detected the transfection efficiency of wtp53 in RB models and explored the feasibility of transfection in vivo RB that was mediated by ultrasound-microbubble.Methods:1.HXO-Rb44 retinoblastoma human cells were grown under the conventional conditions.Cells were pelleted at 1000rpm,and rinsed with Hank at logarithmic phase.After pelleting,cells were suspended in RPMI-1640 medium free fetal calf serum heat inactivated at a concentration of 3.5×107-5.5×107cells/ml.2.12 BALB/C(nu/nu)mice were used randomly. The 5ul suspension of HOX-Rb44 cells were injected into each vitreous cavity.The animals were examined clinically at regular intervals using direct and indirect ophthalmoscopy per 2 days after the injection,to monitor the growth and condition of the grafts as well as the host eye.3.Transfected pC53-SN3 plasmid into nude mice that successful models according to experimental group.Animals were killed 7 days after transfection,removed eyeballs to observed histological features and detected the transfection efficiency of wtp53 gene by RT-PCR.Results:1. At the HOX-Rb44 cells transplantion experiment by vitreous cavity injection, 8 mice , 16 eyes successful In 12 BALB/C(nu/nu)nude mice of random selection.2.At the condition:intensity ultrasound 0.5W/cm2,exposure time 60s,microbubble concentrations of 10%,will not cause injury eye of mice.3.Preliminary findings of the study showed that ultrasound microbubbles can promote the transfection and expression of wtp53 gene in RB tumor tissue.Conclusions:Under certain conditions,ultrasonic microbubbles can promote the transfectiong and expression of wtp53 gene into nude mice xenografts of human RB, and provide a new ideas for gene therapy studies of retinoblastoma.