大鼠神经干细胞通过Wnt/β-catenin途径抑制神经胶质瘤生长  

Rat neural stem cells inhibit C6 glioma cell proliferation through Wnt/β-catenin pathway

作　　者：李一明[1] 李维卿[2] 卢亦成[1] 余宏宇[2] 韩焕兴[3] 何金[2] 徐毅[2] 

机构地区：[1]第二军医大学长征医院神经外科,上海200003 [2]第二军医大学长征医院病理科,上海200003 [3]第二军医大学长征医院实验诊断科,上海200003

出　　处：《第二军医大学学报》2010年第12期1277-1281,共5页Academic Journal of Second Military Medical University

基　　金：国家高技术研究发展计划("863"计划;2005AA001070)~~

摘　　要：目的探讨大鼠神经干细胞对C6成胶质细胞瘤细胞生长的影响,并探讨可能的作用机制。方法采用0.4μm孔径的Transwell小室将C6成胶质细胞瘤细胞和大鼠神经干细胞(NSCs)按不同比例[(C6:NSCs):5:1(2×105:4×104)、1:1(2×105:2×105)、1:5(4×104:2×105)]在无血清培养基中共培养7d作为实验组,以单独培养的C6成胶质细胞瘤细胞作为对照组。采用SCID荷瘤动物模型观察共培养体系中C6成胶质细胞瘤细胞的成瘤能力;利用半定量RT-PCR及蛋白质印迹等方法分析在共培养体系中C6成胶质细胞瘤细胞凋亡相关基因(BMP2、c-Myc、Bcl-2、p53mRNA)及Wnt信号分子蛋白(β-catenin、survivin)的表达。结果与实验组相比,对照组的组织切片中肿瘤细胞恶性程度高,有较多的核分裂像,核/质比例高;大片区域出现单核或多核瘤巨细胞。在实验组中,随着共培养体系中起始神经干细胞比例增高,肿瘤细胞异型性逐步降低。随着共培养体系中起始神经干细胞比例增高,C6成胶质细胞瘤细胞p53mRNA的表达水平逐步增高,BMP2、c-Myc、Bcl-2mRNA表达水平则逐步降低(P<0.05),β-catenin、survivin蛋白表达水平逐步降低(P<0.05)。结论大鼠神经干细胞在胶质瘤微环境中可能通过Wnt/β-catenin途径促进神经胶质瘤细胞凋亡进而抑制神经胶质瘤生长。Objective To study the effect of rat neural stem cells on C6 glioma cell proliferation and to explore the possible mechanism.Methods We co-cultured neural stem cells with C6 glioma cells at different ratios （11[2×10^5 2×10^5],15[4×10^42×10^5],51 [2×10^54×10^4]） in serum-free medium using Transwell chamber culture system for 7 days,and C6 glioma cells cultured alone served as controls.The tumorigenic ability of C6 glioma cells was observed by means of SCID mice transplantation.RT-PCR and Western blotting analysis were used to examine the expression of apoptosis-related proteins （BMP2,c-Myc,Bcl-2 and p53 mRNA） and Wnt signal molecules （β-catenin and survivin）.Results Compared with the experimental group,the tumor cells in the tissue section of the control group had a higher malignant degree,with more mitoses,higher nuclear/cytoplasm ratio,and there were abundant single-or multi-core giant tumor cells in the tissue section.With the increase of neural stem cell proportion in the co-culture system,the tumor cell atypia degree decreased gradually,the expression of p53 mRNA in C6 glioma cells increased gradually,the expression of bone morphogenetic protein 2,Bcl-2,and c-Myc mRNA was decreased significantly （P〈0.05）,and the expression of β-catenin,survivin protein was decreased significantly（P〈0.05）.Conclusion Rat neural stem cells may inhibit the tumorigenicity of C6 glioma cells by promoting apoptosis through Wnt/β-catenin pathway.

关 键 词：神经干细胞 胶质母细胞瘤 WNT/Β-CATENIN信号转导通路 细胞凋亡 

分 类 号：R730.264[医药卫生—肿瘤]