Neuroepithelial stem cell marker nestin regulates the migration, invasion and growth of human gliomas
Neuroepithelial stem cell marker nestin regulates the migration, invasion and growth of human gliomas
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DOI：
10.3892/or.2011.1267
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发表时间：
2011-07-01
期刊：
影响因子：
4.2
通讯作者：
Naito, Zenya
中科院分区：
文献类型：
--
作者：
Ishiwata, Toshiyuki;Teduka, Kiyoshi;Naito, Zenya
巢蛋白是一种 VI 类中间丝蛋白，最初被描述为中枢神经系统发育过程中的神经元干细胞标记物。 Nestin在胶质瘤中表达，其在WHO组织病理学分类等级高的胶质瘤中的表达水平高于低等级的胶质瘤。在本研究中，我们检查了巢蛋白是否调节人神经胶质瘤细胞的生物活性。通过免疫组织化学方法，检查了 10 名人类胶质母细胞瘤患者的巢蛋白表达模式。使用实时 PCR、蛋白质印迹和免疫荧光分析检查了人高级神经胶质瘤细胞系 A172 和人低级神经胶质瘤细胞系 KG-I-C 中巢蛋白的表达水平。将携带靶向巢蛋白的短发夹RNA的表达载体稳定转染至A172（Sh）细胞中。研究了Sh细胞中巢蛋白表达水平降低对细胞生长、迁移、侵袭、细胞外基质粘附以及三维培养板上纤维状肌动蛋白表达的影响。将巢蛋白表达载体瞬时转染至KG-1-C（Nes）细胞中，并检查巢蛋白过表达对细胞生长和迁移的影响。在所有检查的病例中，巢蛋白均在胶质母细胞瘤细胞的细胞质中表达。 Sh细胞的nestin mRNA和蛋白表达水平明显降低，且Sh细胞的生长速度低于假(Sc)细胞。相反，Sh细胞对I型和IV型胶原、纤连蛋白和层粘连蛋白的粘附活性高于Sc细胞。在三维培养板上附着位点的 Sh 细胞集落外围清楚地检测到纤维状肌动蛋白。与Sc细胞相比，Sh细胞的迁移和侵袭受到明显抑制。相反，瞬时转染巢蛋白表达载体的Nes细胞中巢蛋白表达水平显着增加。 Nes细胞的生长速率和运动能力高于模拟细胞。总之，巢蛋白在神经胶质瘤细胞的细胞生长、迁移、侵袭和细胞外基质粘附中发挥重要作用。 Nestin 可能作为神经胶质瘤（包括胶质母细胞瘤）分子靶向治疗的新候选药物。
Nestin, a class VI intermediate filament protein, was originally described as a neuronal stem cell marker during central nervous system development. Nestin is expressed in gliomas, and its expression levels are higher in gliomas with high WHO histopathological classification grades than in those with low grades. In the present study, we examined whether nestin regulates the biological activities of human glioma cells. Immunohistochemically, the nestin expression patterns in 10 human glioblastoma patients were examined. The expression levels of nestin in A172, a human high-grade glioma cell line, and KG-I-C, a human low-grade glioma cell line, were examined using real-time PCR, Western blot and immunofluorescence analyses. An expression vector carrying a short hairpin RNA targeting nestin was stably transfected into A172 (Sh) cells. The effects of decreased expression levels of nestin in Sh cells on cell growth, migration, invasion, adhesion to extracellular matrices and fibrillar actin expression on three-dimensional culture plates were examined. The nestin expression vector was transiently transfected into KG-1-C (Nes) cells, and the effects of the nestin overexpression on cell growth and migration were examined. Nestin was expressed in the cytoplasm of the glioblastoma cells in all cases examined. Sh cells showed marked decreases in the expression levels of nestin mRNA and protein, and the growth rate of Sh cells was lower than that of sham (Sc) cells. In contrast, the adhesion activity of Sh cells to types I and IV collagens, fibronectin and laminin was higher than that of Sc cells. Fibrillar actin was clearly detected at the periphery of colonies of Sh cells at the attachment sites on three-dimensional culture plates. The migration and invasion of Sh cells were markedly inhibited compared with those of Sc cells. In contrast, the levels of nestin expression markedly increased in the Nes cells, which were transiently transfected with the nestin expression vector. The growth rate and motility of Nes cells were higher than those of the mock cells. In conclusion, nestin plays important roles in cell growth, migration, invasion and adhesion to extra-cellular matrices in glioma cells. Nestin may serve as a novel candidate for molecular-targeted therapy for gliomas, including glioblastomas.