trkA基因表达在神经母细胞瘤合成及分泌血管内皮生长因子中作用的研究

Inhibitory Effects of trkA Gene on Synthesis and Secretion of Vascular Endothelial Growth Factor in Human Neuroblastoma Cells

目的 :观察trkA基因在神经母细胞瘤 (NB)中表达对NB合成、分泌血管内皮生长因子 (VEGF)的影响。方法 :采用脂质体转染法建立trkA基因高表达的NB细胞系 ;RT PCR、免疫细胞化学及ELISA技术检测转染前后细胞中VEGF量的变化。结果 :trkA转染后细胞中的VEGFmRNA及VEGF蛋白表达较亲代SY5Y细胞均明显下降 (P均 <0 0 1) ;细胞培养上清液中的VEGF含量也明显下降 (P <0 0 1)。结论 :trkA基因的高表达有效抑制NB细胞合成、分泌VEGF 。

Objective: Our aim was to investigate the role of tyrosine kinase A (trkA) gene inhibiting the synthesis and secretion of vascular endothelial growth factor (VEGF) in human neuroblastoma(NB) cells. Methods: We established TrkA -SY5Y NB cell line which expressing high-level trkA gene with lipofect method. The expressions of VEGFmRNA and VEGF protein in parental SY5Y NB cell line and TrkA -SY5Y NB cell line were detected by using RT-PCR and immuno-cell-chemistry. The quantity of VEGF in cell culture supernatants was determined with ELISA assay. Results: The expressions of VEGF mRNA and VEGF protein were greatly decreased because of the high-level expression of trkA gene in TrkA-SY5Y cells(P<0.01). The quantity of VEGF in cell culture supernatants of TrkA-SY5Y cells was much more lower than that of parent SY5Y(P<0.01). Conclusion: The synthesis and secretion of VEGF in human neuroblastoma cells can be effectively inhibited by high-level expression of trkA gene, which may provides a new way for neuroblastoma angiostatic therapy.