NOS2 expression in glioma cell lines and glioma primary cell cultures: correlation with neurosphere generation and SOX-2 expression
NOS2 expression in glioma cell lines and glioma primary cell cultures: correlation with neurosphere generation and SOX-2 expression
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DOI：
10.18632/oncotarget.16106
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发表时间：
2017-04-11
期刊：
影响因子：
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通讯作者：
Cifone, Maria Grazia
中科院分区：
文献类型：
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作者：
Palumbo, Paola;Miconi, Gianfranca;Cifone, Maria Grazia
一氧化氮与胶质母细胞瘤（GBM）的生物学和进展有关，能够根据细胞暴露的浓度和持续时间影响细胞信号。已提出NOS 2（诱导型一氧化氮合酶）作为几种肿瘤的分子谱的组分，包括神经胶质瘤，神经胶质瘤是最具侵袭性的原发性脑肿瘤之一，其特征在于负责增强对治疗的抗性和肿瘤复发的局部癌症干细胞。在这里，我们研究了NOS 2 mRNA的表达，通过逆转录-PCR在人脑胶质瘤原代培养在几个级别的恶性和胶质瘤干细胞（GSC）衍生的神经球。神经胶质瘤细胞系在干性标志物表达和产生神经球的能力方面均用作阳性对照。在细胞系和原代培养物中检测到基础水平的NOS 2表达，并且在保持在神经球的特定培养基中的培养物中出现显著上调。还对所有细胞培养物进行了免疫荧光分析，以评估SOX-2（GBM中异常上调的干性标志物）的水平。还验证了NOS 2表达和产生神经球的能力之间以及NOS 2和SOX-2水平之间的潜在相关性。结果表明，在所有能产生神经球的原代培养物中检测到较高的NOS 2表达。已经观察到在标准条件下维持的所有细胞培养物中NOS 2表达和SOX-2阳性细胞（%）之间的高度和显著相关性。这些结果揭示了NOS 2作为胶质瘤恶性程度和复发的预后因子的潜在相关性。
Nitric oxide has been implicated in biology and progression of glioblastoma (GBM) being able to influence the cellular signal depending on the concentration and duration of cell exposure. NOS2 (inducible nitric oxide synthase) have been proposed as a component of molecular profile of several tumors, including glioma, one of the most aggressive primary brain tumor featuring local cancer stem cells responsible for enhanced resistance to therapies and for tumor recurrence. Here, we investigated the NOS2 mRNA expression by reverse transcription-PCR in human glioma primary cultures at several grade of malignancy and glioma stem cell (GSC) derived neurospheres. Glioma cell lines were used as positive controls both in terms of stemness marker expression that of capacity of generating neurospheres. NOS2 expression was detected at basal levels in cell lines and primary cultures and appeared significantly up-regulated in cultures kept in the specific medium for neurospheres. The immunofluorescence analysis of all cell cultures to evaluate the levels of SOX-2, a stemness marker aberrantly up-regulated in GBM, was also performed. The potential correlation between NOS2 expression and ability to generate neurospheres and between NOS2 and SOX-2 levels was also verified. The results show that the higher NOS2 expression is detected in all primary cultures able to arise neurosphere. A high and significant correlation between NOS2 expression and SOX-2 positive cells (%) in all cell cultures maintained in standard conditions has been observed. The results shed light on the potential relevance of NOS2 as a prognostic factor for glioma malignancy and recurrence.