论文部分内容阅读
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 investigate the effects of rat neural stem cells on the growth of C6 glioma cells and to explore the possible mechanism. Methods C6 glioma cells and rat neural stem cells (NSCs) were transfected with 0.4% (5: 1) (2 × 105: 4 × 104) (2 × 105: 2 × 105), 1: 5 (4 × 104: 2 × 105)] were cultured in serum-free medium for 7 days as the experimental group and cultured C6 astrocytoma cells alone as the control group. The tumorigenicity of C6 glioblastoma cells in co-culture system was observed by SCID tumor-bearing animal model. The apoptosis of C6 glioblastoma cells in co-culture system was analyzed by semi-quantitative RT-PCR and Western blotting (BMP2, c-Myc, Bcl-2, p53 mRNA) and Wnt signaling protein (β-catenin, survivin) Results Compared with the experimental group, the tumor cells in the control group had high degree of malignancy, more mitosis and high ratio of nuclear / cytosols, and mononuclear or multinucleated giant cells in the large area. In the experimental group, as the percentage of initial neural stem cells in the co-culture system increased, the atypia of the tumor cells gradually decreased. The expression of p53 mRNA in C6 glioblastoma cells gradually increased, while the expression of BMP2, c-Myc and Bcl-2 mRNA gradually decreased (P <0.05), while β-catenin , The expression of survivin protein decreased gradually (P <0.05). Conclusion The neural stem cells of rat may promote glioma cell apoptosis and then inhibit the growth of glioma via the Wnt / β-catenin pathway in glioma microenvironment.