超声微泡介导野生型P53基因转染鼠视网膜母细胞瘤的实验研究

Experimental study of Wild-type p53 gene into transfected the retinoblastoma of nude mices by ultrasound microbubble intensifier

目的:探讨超声微泡造影剂在一定强度的超声波照射下介导野生型p53(wtp53)基因转染荷瘤小鼠的可行性,为实现外源基因高效,定向的转移目的奠定基础。方法:将12只BAlB/C(nu/nu)裸小鼠双眼玻璃体腔接种HXO-Rb44细胞,造模成功后,将动物随机分为2组,第1组于尾静脉注入含质粒的微泡造影剂;第2组尾静脉注射质粒与微泡的混合液,并立即以0.5W/cm2〔1〕的超声波辐照小鼠眼球60s,工作时间控制为20%。转染7天后,处死动物,摘除眼球,RT-PCR检测wtp53基因的表达情况。结果:超声照射组动物的肿瘤组织中均检测到wtp53的mRNA表达,而未照射组动物肿瘤组织中未检测到mRNA的表达。结论:超声微泡能使外源基因wtp53高效的转染小鼠RB肿瘤组织。

Objective：To investigate the feasibility of Wild - type p53 gene transfected into nude mice with retinoblastoma by microbubbles and ultrasound with a certain intensity, and to afford a base for effective and directional gene delivery. Methods：Twelve BAIB/C （nu/nu） nude mice were transplanted with human RB cell - HXO - Rb44 into eyes of the vitreous cavity. After the animal modle done successfully, all animals were randomly divided into two groups, group 1 ： Microbuble contrast agent containing plasmid was injected into nude mice via the tail vein;group 2：the mixture of microbubbles with plasmid was injected as the group 1, and eyeballs were exposed to ultrasound with intensity of 0.5W/cm^2 immediately, and the time were all 60s that working time control at 20%. The animals were sacrificed in the seven day after transfection, and removed the eyeballs, the expression of wtp53 gene was detection by RT- PCR. Results：In the group exposed to ultrasonic, wtp53 mRNA expression were detected in tumor tissue in all animals, but the mRNA expression was not detected in the group without ultrasonic irradiation. Conclusion： Ultrasound microbubbles enable exogenous gene wtp53 transfected into nude mice RB tumor tissues effectively.