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胶质细胞瘤

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PTEN mainly expressedin neurons in our experiment and the absence of PTEN expression in glia may be relatewith the development of gliomas.

PTEN可能主要在神经元中表达,其在胶质细胞的缺失可能和神经胶质瘤的发生发展有关。

Methods The cells were divided into 5 treatment groups(10,25,50,75 and 100 μmol·L-1 QUE), blank control and menstruum control group. The rat C6 cells were cultivated to 1×106·mL-1 in the RPMI 1640 medium, then added into 96 holes board with various doses of QUE by 3 holes per group,and MTT assay was used to observe the proliferation of the cells treated for 24,48 and 72 h. The change of cell cycle was also observed by flow cytometry after the cells were treated with 50 and 100 μmol·L-1 QUE for 48 h. The changes of the protein P53 and Bcl-2 of C6 cells treated with 50 μmol·L-1 QUE for 48 h were detected by immunocytochemical methods.

按QUE浓度分成10、25、50、75及100 μmol·L-15个处理组和空白对照组及溶剂对照组,大鼠脑胶质瘤C6细胞在RPMI 1640培养基中生长达1×106·mL-1后,在96孔板中分别加入上述浓度的QUE继续培养,每组设3复孔,作用24、48及72 h,采用MTT比色法检测QUE对大鼠脑胶质瘤C6细胞的增殖抑制情况,流式细胞术对50及100 μmol·L-1的QUE作用48 h的大鼠脑胶质瘤C6细胞进行周期分析,免疫组化法检测50 μmol·L-1的QUE作用48 h 的p53和bcl-2基因产物。

The pcDNA〓-apoE〓 plasmid was transfected to SK-N-SH Neuroblastoma cell by liposome. It could be screened by G〓. The immunohistochemical assay was shown that neuroblastoma cell transfected by pcDNA〓-apoE〓 expressed the recombinant apoE protein. The intracellular expressed recombinant protein could inhibit the death of neuroblastoma cell induced by beta amyloidal peptides 25-35 fragment.

用pcDNA〓—apoE〓真核表达载体与脂质体共转染神经成纤维胶质瘤细胞,用G〓选择压力筛选,细胞免疫荧光检测表明,pcDNA〓—apoE〓转染的神经成纤维胶质瘤细胞表达了apoE〓重组蛋白,这种内源性的apoE〓重组蛋白对25μM Aβ25—35多肽诱导的神经成纤维胶质瘤细胞的死亡具有拮抗作用。

Against glioma in support of the central nervous system of glial cells.

神经胶质瘤侵害支持中枢神经系统的神经胶质细胞。

At histopathologic analysis, neuronal tumors are usually classified as pure neuronal cell tumors (gangliocytoma, Lhermitte-Duclos disease , central neurocytoma) and mixed neuronal-glial tumors (ganglioglioma, desmoplastic infantile ganglioglioma, dysembryoplastic neuroepithelial tumor, ganglioneuroma).

根据组织病理学的分析,神经元肿瘤通常被分为:单纯的神经元细胞肿瘤(神经节细胞瘤,Lhermitte-Duclos 病〔发育不良性小脑神经节细胞〕瘤),中枢神经细胞瘤)和混合性的神经元-胶质肿瘤(神经节神经胶质瘤,成结缔组织性婴儿节细胞胶质瘤,胚胎发育不良性神经上皮瘤,神经节瘤)。

Methods DCs were prepared from peripherad blood mononuclear induce d with gra-nulocyt e/macrophage colony-stimulating factor and interleukin-4. Apoptosis of glioma cells were induced with γ-radiation. We design these experiment groups including (1) coculture of DCs and apoptotic glioma cells and T cells,(2) coculture of DCs and U937 cells and T cells,(3) coculture of DCs and cultured glioma cell and T cells,(4) coculture of Des and T cell.

用粒-巨噬细胞集落刺激因子加白介素-4(IL-4)从人外周血分化、诱导DCs、γ-射线在体外诱导培养的人脑胶质瘤细胞凋亡,将DCs、T淋巴细胞和凋亡胶质瘤细胞共培养,同时设计不同类型肿瘤细胞(U937及培养胶质瘤细胞)作对照,分离、富集DCs、T淋巴细胞进行免疫应答及肿瘤细胞杀伤试验。

By culturing human brain microvascular endothelial cells and glioma cells separatedly and simutaneously ,and then analyzing their pre- and post-cultune FasL levels, we study the effect of the grouth of glioma cells upon FasL level of human brain microvascular endothelial cells.

我们通过将人脑毛细血管内皮细胞与胶质瘤细胞分别培养及共同培养,对培养前后人脑毛细血管内皮细胞与胶质瘤细胞FasL表达水平的分析,来探讨胶质瘤细胞的生长对人脑毛细血管内皮细胞FasL表达的影响。

Methods At first, retrovirus vectors encoding IFN-γ or IL-4 gene were constructed. They were transfected into PA317 packaging cells by lipofectamine, PA317IFN-γ and PA317IL-4 cells were obtained. C6 glioma cells were infected with replication deficiency retrovius containing IFN-γ or IL-4 gene, cell morphology、cell growth curve and cloning efficiency assay were examined. The intracranial C6 glioma animal model was established in immunocompetent Wistar rats. PA317IFN-γ and PA317IL-4 cells were stereotactically implanted into the tumor areas alone or together. The survival of tumor bearing rats were examined, tumor volumes were measured and immunohistochemical analyses of CD4〓 and CD8〓 T lymphocyte infiltration were performed.

构建和鉴定携带IL-4或IFN-γ基因的逆转录病毒载体,将其导入逆转录病毒包装细胞PA317,通过G418抗性筛选,得到携带目的基因的包装细胞并鉴定;应用携带IL-4或IFN-γ基因的复制缺陷型逆转录病毒感染C6胶质瘤细胞,观察对C6胶质瘤细胞形态、细胞生长曲线和克隆形成率的影响;建立C6胶质瘤大鼠脑内移植动物模型,将携带IL-4和IFN-γ基因的逆转录病毒包装细胞分别或联合注射到脑内荷瘤大鼠的肿瘤组织中,观察其治疗作用,并初步探讨其机制。

In Part 1 of our research, BGSCs were sorted through immunomagnetic beads marking by CD133 and cultured in vitro, and character as a stem cell was identified by stem cell markers (CD133 and Nestin) and differentiated cell markers [ microtubule-associated protein 2(MAP2), glial acidic fibrillary protein and myelin basic protein] , ultrastructure observing with electron microscopeand engrafting to severe combined immunodeficiency mice for tumorigenesis test. The results were as following: Only a small subset of CD133+ glioma cells in glioma cell lines and fresh specimens from various pathologic grade could express stem cell markers CD133 and Nestin, view ultrastructure of a stem cell and be capacity of serial passage in culture. These CD133+ cells possese a marked capacity for multipotent differentiation and could differentiate into tumor cells expressing MAP2,β-TubulinⅢ, GFAP and MBP; When engrafted into SCID mice, they can generate and form tumors that phenotypically resembl the tumor from the patient.

在本课题中,在第一部分实验中采用以CD133为标志的免疫磁珠法从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞并进行体外培养,通过免疫荧光技术检测干细胞标志物CD133、Nestin,诱导分化后检查分化细胞标志物MAP2、GFAP、MBP以及电镜超微结构观察和移植SCID鼠致瘤实验,对其干细胞特性加以鉴定,得到如下结果:不同病理分级的新鲜胶质瘤标本和胶质瘤细胞株中存在一小部分CD133+的胶质瘤细胞,能表达干细胞的标志物CD133和Nestin,符合干细胞的超微结构特点,体外培养能连续传代;具有多向分化潜能:诱导分化后能产生MAP2、β-TubulinⅢ、GFAP、MBP染色阳性的细胞;移植SCID鼠后能形成与亲本肿瘤表型一致的移植瘤。

Neural stem cells have a strong self-renew mechanism and it can transform after a little break. Neural stem cells have a long term survival, which mean that it has more probability of wrong copy than mature cells. These cells are formed glioma stem cells in the end. The genes who adjust neural stem cells can express in glioma stem cells, which hold out glioma stem cells from neural stem cells. There is another presume that glioma stem cells come from differentiated cells. Through the gene break of these cells, they can obtain characteristics of stem cells, then form glioma stem cells.

神经干细胞具有很强的自我更新机制,获得较少突变即有可能恶性转化,而且干细胞存活时间较长,这意味着干细胞比成熟细胞发生细胞复制的错误几率更大,因外界环境的刺激而发生突变的机会更多,最终形成脑胶质瘤干细胞,同时调节神经干细胞增殖和自我更新的基因在脑胶质瘤的脑胶质瘤干细胞中也表达,这也是支持神经干细胞是脑胶质瘤干细胞来源的;也有推测认为它可能起源于已分化的细胞,由这些细胞突变发生去分化得来,并通过基因突变而获得了干细胞自我更新的特性,从而形成脑胶质瘤干细胞。

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