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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
Objective TrkA gene is one of the markers of neuroblastoma (NB) with good prognosis. High expression of TrkA gene may not only inhibit NB proliferation and induce benign differentiation, but may also affect the growth of NB. This experiment explored the role of the TrkA gene in NB angiogenesis. Methods Fifteen nude mice were randomly divided into control group, empty vector group and experimental group (5 in each group). The TrkA gene and pBPSTR1 empty vector were transfected into NBSY5Y cells by lipofection method, and named as SY5Y TrkA and SY5Y Vec cells respectively. The untransfected cells were SY5Y cells. The SY5Y, SY5Y, Vec and SY5Y TrkA cells were respectively inoculated subcutaneously in the control group, the empty vector group and the experimental group, and the animals were sacrificed 50 days after the inoculation. The tumors were excised and the tumor volume was measured. The expression of tumor markers was detected by RT-PCR and immunohistochemistry Expression of vascular endothelial growth factor (VEGF); calculate microvessel density (MVD). Results TrkA expression in SY5Y TrkA cells was significantly higher than that in SY5Y Vec and SY5Y cells groups (P <0.01). The final tumor volume in experimental group was smaller than that in control group and empty vector group (0.39 ± 0.02cm3 vs 1.74 ± (0.39 ± 0.22cm3 vs1.80 ± 0.75cm3), the difference was significant (all P <0.01); The expression of VEGFmRNA in experimental group was lower than that in control group and empty vector (0 .1 6 ± 0 .0 9 vs 1 .4 5 ± 0 .77); (0 .1 6 ± 0 .0 9 vs 1 .35 ± 0 .71), the difference was significant (all P <0. 0 1). The expression of VEGF protein in the experimental group was lower than that in the control group and the empty vector group (2.000 ± 0.600 vs 5.67 ± 0.49); (2.00 ± 0.60v