Human cortical glial tumors contain neural stem-like cells expressing astroglial and neuronal markers in vitro
Human cortical glial tumors contain neural stem-like cells expressing astroglial and neuronal markers in vitro
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DOI：
10.1002/glia.10094
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发表时间：
2002-09-01
期刊：
影响因子：
6.2
通讯作者：
Steindler, DA
中科院分区：
文献类型：
--
作者：
Ignatova, TN;Kukekov, VG;Steindler, DA
来自哺乳动物CNS的神经源性区域的神经干细胞在体外培养系统中克隆形成，所述体外培养系统利用在补充有甲基纤维素和多效性生长因子EGF、FGF 2和胰岛素的培养基中的血清和锚定撤回。本研究的目的是测试是否皮质神经胶质瘤包含干细胞样细胞能够，在这种培养系统下，形成克隆表现出克隆内异质性的神经谱系特异性蛋白质的表达。在p53突变的临床肿瘤标本和正常人中枢神经系统的神经源性区域中，克隆形成细胞的频率很高（约0.1-10 × 10 ~（-3）），这表明在这种培养系统中形成克隆的能力是由表观遗传学诱导的。RT-PCR分析人群的正常脑和肿瘤来源的姐妹克隆巢蛋白，神经元特异性烯醇化酶，和胶质细胞酸性蛋白（GFAP）的成绩单。然而，肿瘤衍生的克隆是不同的克隆来自正常大脑的神经原性区域的转录本的表达与神经细胞的命运决定相关的基因通过刻缺蛋白信号通路（德尔塔和锯齿状），并在G2或有丝分裂期（生存素）的细胞存活。此外，个别神经胶质瘤衍生的克隆含有分别对GFAP或神经元β-III微管蛋白呈免疫阳性的细胞，以及共表达神经胶质和神经元标记物两者的单细胞。这些数据表明，潜在的关键干细胞的特性可以表观遗传诱导的生长条件，不仅在正常中枢神经系统的神经原性区域的细胞，但也在皮质神经胶质肿瘤的细胞。此外，对表观遗传刺激具有遗传缺陷反应的肿瘤干细胞样细胞可能有助于神经胶质瘤的发生和神经胶质瘤的发育病理异质性。
Neural stem cells from neurogenic regions of mammalian CNS are clonegenic in an in vitro culture system exploiting serum and anchorage withdrawal in medium supplemented with methyl cellulose and the pleiotropic growth factors EGF, FGF2, and insulin. The aim of this study was to test whether cortical glial tumors contain stem-like cells capable, under this culture system, of forming clones showing intraclonal heterogeneity in the expression of neural lineage-specific proteins. The high frequencies of clone-forming cells (about 0.1-10 x 10(-3)) in clinical tumor specimens with mutated p53, and in neurogenic regions of normal human CNS, suggest that the ability to form clones in this culture system is induced epigenetically. RT-PCR analyses of populations of normal brain- and tumor-derived sister clones revealed transcripts for nestin, neuron-specific enolase, and glial fibrillary acidic protein (GFAP). However, the tumor-derived clones were different from clones derived from neurogenic regions of normal brain in the expression of transcripts specific for genes associated with neural cell fate determination via the Notch-signaling pathway (Delta and Jagged), and cell survival at G2 or mitotic phases (Survivin). Moreover, the individual glioma-derived clones contain cells immunopositive separately for GFAP or neuronal beta-III tubulin, as well as single cells coexpressing both glial and neuronal markers. The data suggest that the latent critical stem cell characteristics can be epigenetically induced by growth conditions not only in cells from neurogenic regions of normal CNS but also in cells from cortical glial tumors. Moreover, tumor stem-like cells with genetically defective responses to epigenetic stimuli may contribute to gliomagenesis and the developmental pathological heterogeneity of glial tumors.