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神经纤毛蛋白-1是胶质母细胞瘤中一种胶质细胞系源性神经营养因子受体。

Neuropilin-1 is a glial cell line-derived neurotrophic factor receptor in glioblastoma.

作者信息

Sun Shen, Lei Yu, Li Qi, Wu Yue, Zhang Lin, Mu Pei-Pei, Ji Guang-Quan, Tang Chuan-Xi, Wang Yu-Qian, Gao Jian, Gao Jin, Li Li, Zhuo Lang, Li Yun-Qing, Gao Dian-Shuai

机构信息

Department of Anatomy and Histology, The Fourth Military Medical University, Xi'an, Shanxi, China.

Department of Neurobiology and Anatomy, Xuzhou Key Laboratory of Neurobiology, Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, Jiangsu, China.

出版信息

Oncotarget. 2017 Jun 27;8(43):74019-74035. doi: 10.18632/oncotarget.18630. eCollection 2017 Sep 26.

DOI:10.18632/oncotarget.18630
PMID:29088765
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5650320/
Abstract

The aim of this study was to identify the receptor for glial cell line-derived neurotrophic factor (GDNF) in glioblastoma multiforme (GBM). After GST pull-down assays, membrane proteins purified from C6 rat glioma cells were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS). The differentially expressed proteins were annotated using Gene Ontology, and neuropilin-1 (NRP1) was identified as the putative GDNF receptor in glioma. NRP1 was more highly expressed in human GBM brains and C6 rat glioma cells than in normal human brains or primary rat astrocytes. Immunofluorescence staining showed that NRP1 was recruited to the membrane by GDNF, and NRP1 co-immunoprecipitated with GDNF. Using the NRP1 and GDNF protein structures to assess molecular docking in the ZDOCK server and visualization with the PyMOL Molecular Graphics System revealed 8 H-bonds and stable positive and negative electrostatic interactions between NRP1 and GDNF. RNAi knockdown of NRP1 reduced proliferation of C6 glioma cells when stimulated with GDNF. NRP1 was an independent risk factor for both survival and recurrence in GBM patients. High NRP1 mRNA expression correlated with shorter OS and DFS (OS: χ=4.6720, =0.0307; DFS: χ=11.013, =0.0009). NRP1 is thus a GDNF receptor in glioma cells and a potential therapeutic target.

摘要

本研究的目的是鉴定多形性胶质母细胞瘤（GBM）中胶质细胞系源性神经营养因子（GDNF）的受体。在进行谷胱甘肽S-转移酶（GST）下拉分析后，对从C6大鼠胶质瘤细胞中纯化的膜蛋白进行液相色谱-串联质谱分析（LC-MS/MS）。使用基因本体论对差异表达的蛋白质进行注释，并将神经纤毛蛋白-1（NRP1）鉴定为胶质瘤中假定的GDNF受体。与正常人脑或原代大鼠星形胶质细胞相比，NRP1在人GBM脑和C6大鼠胶质瘤细胞中表达更高。免疫荧光染色显示，GDNF可将NRP1募集到细胞膜上，且NRP1与GDNF进行了共免疫沉淀。利用NRP1和GDNF的蛋白质结构在ZDOCK服务器中评估分子对接，并使用PyMOL分子图形系统进行可视化分析，结果显示NRP1与GDNF之间存在8个氢键以及稳定的正负静电相互作用。当用GDNF刺激时，RNA干扰敲低NRP1可降低C6胶质瘤细胞的增殖。NRP1是GBM患者生存和复发的独立危险因素。高NRP1 mRNA表达与较短的总生存期（OS）和无病生存期（DFS）相关（OS：χ=4.6720，P=0.0307；DFS：χ=11.013，P=0.0009）。因此，NRP1是胶质瘤细胞中的GDNF受体，也是一个潜在的治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/75f2/5650320/2b45ab18e047/oncotarget-08-74019-g010.jpg
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