神经生长因子受体对神经母细胞瘤血管生成作用的研究(英文)

Role of nerve growth factor receptor in neuroblastoma angiogenesis

摘要:

目的 神经生长因子受体 (TrkA)基因是神经母细胞瘤 (NB)良好预后的标识之一 ,其高表达不仅可以抑制NB增殖和诱导良性分化 ,对NB的血管生长可能有一定的影响。该实验探讨TrkA基因对NB血管生成的作用。方法 1 5只裸鼠随机分为对照组、空载体组和实验组 (每组 5只 )。利用脂质体转染法将TrkA基因和pBPSTR1空载体转入NBSY5Y细胞 ,分别命名为SY5Y TrkA及SY5Y Vec细胞 ,未转染细胞为SY5Y细胞。将SY5Y ,SY5Y Vec ,SY5Y TrkA细胞分别接种在对照组、空载体组和实验组裸鼠皮下 ,接种 5 0d后处死动物 ,切除肿瘤 ,测量肿瘤体积 ;用RT PCR及免疫组织化学技术检测肿瘤内血管内皮生长因子 (VEGF)的表达 ;计算微血管密度 (MVD)。结果 SY5Y TrkA细胞TrkA表达明显高于SY5Y Vec及SY5Y细胞组 (P <0 .0 1 ) ;实验组肿瘤终体积小于对照组及空载体组 (0 .39± 0 .0 2cm3 vs 1 .74± 0 .4 9cm3 ) ;(0 .39± 0 .0 2cm3 vs 1 .80± 0 .75cm3 ) ,差异有显著性 (均P <0 .0 1 ) ;实验组VEGFmRNA表达低于对照组及空载体组 (0 .1 6± 0 .0 9vs 1 .4 5± 0 .77) ;(0 .1 6± 0 .0 9vs 1 .35± 0 .71 ) ,差异有显著性 (均P <0 .0 1 ) ;实验组VEGF蛋白表达低于对照组及空载体组 (2 .0 0± 0 .6 0vs 5 .6 7± 0 .4 9) ;(2 .0 0± 0 .6 0v

Abstract:

Objective The nerve growth factor receptor (TrkA) gene is an independent marker of the favourable prognosis of neuroblastoma (NB). High expression of TrkA can not only induce differentiation and inhitit proliferation of NB cells but can also be involved in the regulation of tumor angiogenesis. This study aims to investigate the role of the TrkA gene in human NB angiogenesis. Methods The NB SY5Y cells were named SY5Y TrkA cells and SY5Y Vec cells after being transfected with TrkA gene and PBPSTR1 empty vector respectively, and the NB SY5Y cells without transfection were named the SY5Y cells. Fifteen nude mice were assigned into a Control group, an Empty Vec group an Experiment group (n=5 each). The SY5Y cells of the Control group, the SY5Y Vec cells of the Empty Vec group and the SY5Y TrkA cells of the Experiment group were inoculated. The mice were then sacrificed 50 days after inoculation. The tumor volume was measured; the microvessel density (MVD) of the tumor was evaluated and the expression of vascular endothelial growth factor (VEGF) was determined by RT PCR and immunohistochemistry. Results The tumor volume in the Experiment group ( 0.39 ± 0.02 cm 3 ) was significantly smaller than that of the Control group ( 1.74 ± 0.49 cm 3 ) and also was smaller than the Empty Vec group ( 1.80 ± 0.75 cm 3 ) (P< 0.01 ). The expressions of VEGF mRNA and VEGF protein in the Experiment group were significantly lower than those of both the Control and the Empty Vec groups (P< 0.01 ). The MVD in the Experiment group ( 4.08 ± 4.72% ) was also significantly lower than that of the Control ( 27.21 ± 14.58% ) and Empty Vec groups ( 27.76 ± 14.15% ) (P< 0.01 ). Conclusions Angiogenesis and tumor growth of human NB can be effectively inhibited by the TrkA gene. This experiment provides a theoretical basis for the treatment of NB with anti angiogenesis gene therapy.