The new prognosis assessment method which the Interaction between NGFR and GM1 is applied to, and NGF sensitive regulatory differentiation derivation therapy for neuroblastma patients

我们的目的是将NGFR （TrkA）与GM1的相互作用应用于神经母细胞瘤的临床诊断和治疗。我们研究了GM1的检测方法，并结合NGFR （TrkA）进行检测，覆盖临床样本。提高免疫印迹法和薄层色谱法的灵敏度。因为不存在抗gm1的高敏抗体。采用霍乱毒素B亚单位（CTB）检测。虽然结合Trk特异性抗体的免疫沉淀法是可行的，但结合Trk的GM1的检测在灵敏度上是困难的。对会话状态变化的临床样本的分析不能适应这一程序。因此，我们通过细胞系集中研究NGFR （TrkA）与GM1相互作用的详细机制。利用共聚焦激光显微镜荧光抗体技术，我们发现NGFR （TrkA）和GM1在可分化为神经元的PC12细胞和神经母细胞瘤细胞株NB-1细胞表面分布情况有显著差异。提示神经母细胞在NGF刺激下不能向神经元分化是由于Trk和GM1的相互作用系统被破坏所致。但是，如果这种机制在分化调控中占主导地位，我们就无法解释高Trk表达的神经母细胞在NGF高浓度培养条件下的分化机制。我们假设在低浓度NGF作用下，高Trk表达细胞发生竞争性抑制机制，并将PC12细胞系与能被NGF分化的PCTrk （Trk高表达PC12）细胞系进行比较。本研究为神经母细胞瘤的新诊断和治疗提供了重要的发现。

our purpose was the application of the interaction with GM1 of NGFR (TrkA) to the clinical diagnosis and the cure of neuroblastoma. We studied the detection method of GM1 which combined in NGFR (TrkA) was examined to cover clinical samples. Increase in sensitivity of the immunoblotting method and the TLC (thin layer chromatography) was tried. Because an anti-GM1 high sensitive antibody didn't exist. It was detected by using CTB (cholera toxin B subunit). Though the immunoprecipitation method with a Trk specific antibody was possible the detection of GM1 which combined in Trk mastered distress in the point of the sensitivity. The analysis of the clinical samples which conversation states varied couldn't adapt this procedure. So, we concentrated the detailed mechanism of NGFR (TrkA) and the GM1 interaction by using cell lines. We found the cell surface distribution situation of NGFR (TrkA) and GM1 was remarkably different in the PC12 cell which can differentiate to neuron, and the neuroblastoma cell line NB-1 by the fluorescent antibody technique which a confocal laser microscope was used for. This result suggested that neuroblastma cannot differentiate to neuron with NGF stimulation because of the destruction of the interaction system between Trk and GM1. But, if this mechanism is dominant for the differentiation regulation, we cannot explain the mechanism which a hyper Trk expression neuroblastma differentiates under the NGF high concentration culture condition. We compared PC12 cell line with the PCTrk (Trk hyper expression PC12) cell line that can differentiate by NGF on the assumption that the competitive inhibition mechanism occurred in the hyper Trk expression cells under the low concentration of NGF. We could get essential findings for new diagnosis and the cure of neuroblastoma in this study.