NOS2 expression in glioma cell lines and glioma primary cell cultures: correlation with neurosphere generation and SOX-2 expression
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Paola Palumbo1, Gianfranca Miconi1, Benedetta Cinque1, Francesca Lombardi1, Cristina La Torre1, Soheila Raysi Dehcordi1,2, Renato Galzio1,2, Annamaria Cimini1,3,4, Antonio Giordano3,5, Maria Grazia Cifone1
1Department of Life, Health and Environmental Sciences, University of L’Aquila, L’Aquila, Italy
2Department of Surgery, Operative Unit of Neurosurgery, San Salvatore Hospital, L’Aquila, Italy
3Sbarro Institute for Cancer Research and Molecular Medicine and Center for Biotechnology, Temple University, Philadelphia, PA, USA
4National Institute for Nuclear Physics (INFN), Gran Sasso National Laboratory (LNGS), Assergi, Italy
5Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy
Paola Palumbo, email: [email protected]
Keywords: glioma, primary cultures, cancer stem cell, NOS2, SOX-2
Received: December 07, 2016 Accepted: February 15, 2017 Published: March 10, 2017
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.
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