Differential Expression and Cellular Distribution of γ-Tubulin and βIII-Tubulin in Medulloblastomas and Human Medulloblastoma Cell Lines
Differential Expression and Cellular Distribution of γ-Tubulin and βIII-Tubulin in Medulloblastomas and Human Medulloblastoma Cell Lines
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DOI：
10.1002/jcp.22077
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发表时间：
2010-05-01
影响因子：
5.6
通讯作者：
Katsetos, Christos D.
中科院分区：
文献类型：
--
作者：
Caracciolo, Valentina;D'Agostino, Luca;Katsetos, Christos D.
在之前的研究中，我们发现γ -微管蛋白和β - iii -微管蛋白在人胶质母细胞瘤和胶质母细胞瘤细胞系中的过表达和异位亚细胞分布（Katsetos et al., 2006,] Neuropathol Exp Neurol 65:455-467; Katsetos et al., 2007, Neurochem Res 32:1387-1398）。我们在手术切除的髓母细胞瘤（n = 20）和人髓母细胞瘤细胞系D283 Med和DAOY中测定了γ -微管蛋白的表达。在临床组织样本中，γ -微管蛋白标记的免疫组织化学分布是普遍的，并且与神经生成呈负相关。γ -微管蛋白的过度表达在低分化、增殖的肿瘤细胞中广泛存在，但在处于神经新生的静止分化的肿瘤细胞中却显著减少，这一点通过β iii -微管蛋白免疫标记得到了强调。通过实时荧光定量PCR，在两种细胞系中均检测到TUBG1、TUBG2和TUBB3基因的γ -微管蛋白转录本，但与人胶质母细胞瘤细胞系相比，表达不明显。免疫印迹显示，在细胞周期的不同阶段，γ -微管蛋白和β iii -微管蛋白的含量相当；然而，与dao细胞相比，D283 Med细胞中检测到更多的γ -微管蛋白。除了预期的中心体相关分布外，间期D283 Med细胞主要表现为弥漫性细胞质γ -微管蛋白定位。在有丝分裂纺锤体中检测到强大的β iii -微管蛋白免疫反应性。我们的数据表明，γ -微管蛋白的过表达可能与髓母细胞瘤的表型去分化（发育不全）和肿瘤进展有关，并可能作为一种有前景的肿瘤标志物。j .细胞。中国生物医学工程学报，2009,31(2):519-529。(C) 2010 Wiley-Liss, Inc。
In previous studies, we have shown overexpression and ectopic subcellular distribution of gamma-tubulin and beta III-tubulin in human glioblastomas and glioblastoma cell lines (Katsetos et al., 2006,] Neuropathol Exp Neurol 65:455-467; Katsetos et al., 2007, Neurochem Res 32:1387-1398). Here we determined the expression of gamma-tubulin in surgically excised medulloblastomas (n = 20) and in the human medulloblastoma cell lines D283 Med and DAOY. In clinical tissue samples, the immunohistochemical distribution of gamma-tubulin labeling was pervasive and inversely related to neuritogenesis. Overexpression of gamma-tubulin was widespread in poorly differentiated, proliferating tumor cells but was significantly diminished in quiescent differentiating tumor cells undergoing neuritogenesis, highlighted by beta III-tubulin immunolabeling. By quantitative real-time PCR, gamma-tubulin transcripts for TUBG1, TUBG2, and TUBB3 genes were detected in both cell lines but expression was less prominent when compared with the human glioblastoma cell lines. Immunoblotting revealed comparable amounts of gamma-tubulin and beta III-tubulin in different phases of cell cycle; however, a larger amount of gamma-tubulin was detected in D283 Med when compared with DAOY cells. Interphase D283 Med cells exhibited predominantly diffuse cytoplasmic gamma-tubulin localization, in addition to the expected centrosome-associated distribution. Robust beta III-tubulin immunoreactivity was detected in mitotic spindles of DAOY cells. Our data indicate that overexpression of gamma-tubulin may be linked to phenotypic dedifferentiation (anaplasia) and tumor progression in medulloblastomas and may potentially serve as a promising tumor marker. J. Cell. Physiol. 223: 519-529,2010. (C) 2010 Wiley-Liss, Inc.