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

1999 Fiscal Year Final Research Report Summary

• Project/Area Number: 10671668
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatric surgery
• Research Institution: Nagoya University
• Principal Investigator: IWATA Hiroyuki Nagoya University, Radioisotope Research Center, Res. Assoc, アイソトープ総合センター, 助手 (30273197)
• Co-Investigator(Kenkyū-buntansha): ISHIGURO Yukio Japanese Red Cross Aichi Juniar College, The Department of Narsing, Prof, 看護学科, 教授 (80142173) ; HAMAGUCHI Michinari School of Medicien, Prof, 医学部, 教授 (90135351)
• Project Period (FY): 1998 – 1999
• Keywords: NGFR / TrkA / GM1 / PC12 / PC12

Research Abstract

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.