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目的探讨载自杀基因的靶向微泡联合超声辐照对视网膜母细胞瘤(RB)的抑制作用。方法制备携带单纯疱疹病毒Ⅰ型胸苷激酶(HSV1-tk)质粒和血管内皮细胞生长因子受体2抗体的靶向微泡,分为空白对照组、细胞+质粒组、细胞+质粒+Sono Vue组、细胞+靶向微泡组、细胞+质粒+超声辐照组、细胞+质粒+Sono Vue+超声辐照组及细胞+靶向微泡+超声辐照组进行实验。荧光显微镜观察各组基因转染情况,流式细胞仪检测转染率,加入丙氧鸟苷(GCV)后检测细胞的抑制率。结果细胞+靶向微泡+超声辐照组的转染率为(24.78±1.04)%,高于细胞+质粒+Sono Vue+超声辐照组(14.31±0.69)%,差异有统计学意义(P<0.05)。随着GCV浓度的增加和培养时间的延长,各组抑制率逐渐升高。当GCV浓度为100 mg/L,培养96 h后,细胞+靶向微泡+超声辐照组对RB细胞的抑制率达(92.91±1.71)%。结论加入GCV后,携带HSV1-tk基因的靶向微泡联合超声能有效地抑制RB细胞。
Objective To investigate the inhibitory effect of suicide gene-containing microbubbles combined with ultrasound on retinoblastoma (RB). Methods The targeted microbubbles carrying herpes simplex virus type 1 thymidine kinase (HSV1-tk) plasmid and vascular endothelial growth factor receptor 2 antibody were prepared and divided into blank control group, cell + plasmid group, cell + plasmid + Sono Vue Group, cell + target microbubble group, cell + plasmid + ultrasonic irradiation group, cell + plasmid + Sono Vue + ultrasonic irradiation group and cell + targeted microbubble + ultrasonic irradiation group. The transfection efficiency of each group was observed by fluorescence microscopy, the transfection efficiency was detected by flow cytometry, and the inhibition rate of cells was detected by adding gavage (GCV). Results The transfection rate of cells + targeted microbubbles + ultrasound irradiation group was (24.78 ± 1.04)%, which was significantly higher than that of cell + plasmid + Sono Vue + irradiation group (14.31 ± 0.69)%, P < <0.05). With the increase of GCV concentration and culture time, the inhibition rate of each group increased gradually. When the GCV concentration was 100 mg / L, the inhibitory rate of RB + cells was (92.91 ± 1.71)% after cells were cultured for 96 h. Conclusion After adding GCV, targeted microbubbles carrying HSV1-tk gene combined with ultrasound can effectively inhibit RB cells.