聚焦超声对斑马鱼神经胶质瘤模型的治疗研究

Effect of focused ultrasound on neuroglioma model of zebrafish

摘要:

目的：通过研究聚焦超声对斑马鱼神经胶质瘤模型治疗后肿瘤组织和肿瘤血管的变化，探讨其对神经胶质瘤及肿瘤血管的影响。方法：将72只转基因斑马鱼（fli：GFP）随机分为3组，每组24只，分别是对照组（Fli组）、肿瘤组（U87组）、治疗组（U87+HIFU组）。在受精2 d后，对肿瘤组和治疗组注射神经胶质瘤细胞（U87-RFP），注射1 d后对治疗组进行聚焦超声治疗。在治疗后连续２ d用荧光显微镜观察３组斑马鱼的肿瘤大小及肿瘤血管增生情况，qPCR法检测血管内皮生长因子（vascular endothelial growth factor，VEGF）Aa和血管内皮生长因子受体（vascular endothelial growth factor receptor，VEGFR）２的mRNA的表达情况。结果：U87组斑马鱼卵黄内有大量神经胶质瘤细胞且肿瘤血管明显增生，VEGF Aa和VEGFR2的mRNA表达升高 （VEGF Aa：Fli：1.01±0.13 vs. U87：1.45±0.10，P=0.002；VEGFR2：Fli：1.01±0.12 vs. U87：1.41±0.07，P=0.001）；经聚焦超声治疗后，斑马鱼卵黄内的神经胶质瘤细胞显著减少，血管增生不明显，VEGF Aa和VEGFR2的mRNA表达也较U87组降低（VEGF Aa：U87+HIFU：1.14±0.05 vs. U87：1.45±0.10，P=0.009；VEGFR2：U87+HIFU：1.16±0.06 vs. U87：1.41±0.07，P=0.012）。结论：聚焦超声能有效杀灭神经胶质瘤细胞，并且能减少肿瘤血管的生成，对治疗神经胶质瘤具有积极作用。

Abstract:

Objective：To explore the treatment effect of focused ultrasound on neuroglioma model of zebrafish. Methods：Transgenic zebrafishes（fli：GFP） were randomly divided into control group，U87 group and U87+HIFU group. Neuroglioma cells（U87-RFP） were injected into U87 group and U87+HIFU group after 2 d fertilization and focused ultrasound treatment was performed in U87+HIFU group after 24 h. The tumor size and the vessels were observed using fluorescence microscopy on the 1st d and the 2nd d after the treatment. The mRNA expression of vascular endothelial growth factor（VEGF） Aa and vascular endothelial growth factor receptor 2 VEGFR2 were evaluated by qPCR. Results：In U87 group，a large number of neuroglioma cells and the newly formed vessels induced by neuroglioma were easily observed in zebrafish’s yolk ball，while after focused ultrasound treatment，neuroglioma cells and the ves－sels were significantly decreased. qPCR suggested that the mRNA of VEGF Aa and VEGFR2 were highly expressed in U87 group（VEGF Aa：Fli：1.01±0.13 vs. U87：1.45±0.10，P=0.002；VEGFR2：Fli：1.01±0.12 vs. U87：1.41±0.07，P=0.001），but after focused ultrasound treatment，the expression of VEGF Aa and VEGFR2 were inhibited（VEGF Aa：U87+HIFU：1.14±0.05 vs. U87：1.45±0.10，P=0.009；VEGFR2：U87+HIFU：1.16±0.06 vs. U87：1.41±0.07，P=0.012）. Conclusion：Focused ultrasound treatment can effectively kill the neuroglioma cells and decrease the tumor angiogenesis，which shows a positive treat effect on neuroglioma.