Department of Life, Health & Environmental Sciences, University of L'Aquila, Building Delta 6, Coppito, 67100 L'Aquila, Italy.
Operative Unit of Neurosurgery, San Salvatore Hospital, 67100 L'Aquila, Italy.
Int J Mol Sci. 2018 Sep 17;19(9):2801. doi: 10.3390/ijms19092801.
Aberrant nitric oxide synthase 2 (NOS2) expression has been suggested as an interesting therapeutic target that is being implicated as a component of the molecular profile of several human malignant tumors, including glioblastoma, which is the most aggressive brain tumor with limited therapeutic options and poor prognosis. The aim of the present work was to evaluate the effect of 1400W, a specific NOS2 inhibitor, on human glioma cells in terms of clonogenic potential, proliferation, migration rate, and neurosphere generation ability. NOS2 expression was determined by Western blotting. Nitric oxide (NO) production was measured through nitrite level determination. The trypan blue exclusion test and the plate colony formation assay were performed to evaluate cell proliferation and clonogenic potential. Cell proliferation and migration ability was assessed by the wound-healing assay. Neurosphere generation in a specific stemcell medium was investigated. NOS2 was confirmed to be expressed in both the glioma cell line and a human glioma primary culture, and overexpressed in relative derived neurospheres. Experiments that aimed to evaluate the influence of 1400W on U-87 MG, T98G (glioblastoma cell lines) and primary glioma cells sustained the crucial role played by NOS2 in proliferation, colony formation, migration, and neurosphere generation, thus supporting the emerging relevance of a NOS2/NO system as a prognostic factor for glioma malignancy and recurrence.
异常的一氧化氮合酶 2（NOS2）表达已被认为是一个有趣的治疗靶点，它被认为是包括神经胶质瘤在内的几种人类恶性肿瘤的分子特征的组成部分，神经胶质瘤是最具侵袭性的脑肿瘤，治疗选择有限，预后不良。本工作的目的是评估特定的 NOS2 抑制剂 1400W 对神经胶质瘤细胞的克隆形成潜力、增殖、迁移率和神经球生成能力的影响。通过 Western blot 测定 NOS2 表达。通过亚硝酸盐水平测定来测量一氧化氮（NO）的产生。通过台盼蓝排除试验和平板集落形成试验评估细胞增殖和克隆形成潜力。通过划痕愈合试验评估细胞增殖和迁移能力。在特定的干细胞培养基中研究神经球生成。NOS2 在神经胶质瘤细胞系和人神经胶质瘤原代培养物中均有表达，在相对衍生的神经球中过表达。旨在评估 1400W 对 U-87 MG、T98G（神经胶质瘤细胞系）和原代神经胶质瘤细胞影响的实验证实了 NOS2 在增殖、集落形成、迁移和神经球生成中的关键作用，从而支持 NOS2/NO 系统作为神经胶质瘤恶性和复发的预后因素的新兴相关性。
