D'Amico Agata Grazia, Maugeri Grazia, Magrì Benedetta, Giunta Salvatore, Saccone Salvatore, Federico Concetta, Pricoco Elisabetta, Broggi Giuseppe, Caltabiano Rosario, Musumeci Giuseppe, Reglodi Dora, D'Agata Velia
Department of Drug and Health Sciences, University of Catania, I‑95125 Catania, Italy.
Department of Biomedical and Biotechnological Sciences, Section of Anatomy, Histology and Movement Sciences, University of Catania, I‑95100 Catania, Italy.
Int J Oncol. 2023 Jan;62(1). doi: 10.3892/ijo.2022.5462. Epub 2022 Dec 9.
Glioblastoma multiforme (GBM) is a brain cancer with a poor prognosis that affects adults. This is a solid tumor characterized by a high rate of cell migration and invasion. The uncontrolled cell proliferation creates hypoxic niches in the tumor mass, which leads to the overexpression of hypoxia‑inducible factors (HIFs). This induces the activation of the vascular endothelial growth factor (VEGF), which is responsible for uncontrolled neoangiogenesis. Recent studies have demonstrated the anti‑invasive effect of pituitary adenylate cyclase‑activating peptide (PACAP) in GBM. PACAP effects on the central nervous system are also mediated through the activity‑dependent neuroprotective protein (ADNP) activation. To date, no evidence exists regarding its role in GBM. Therefore, the ADNP involvement in GBM was investigated. By analyzing ADNP expression in a human GBM sample through confocal microscopy, a high ADNP immunoreactivity was detected in most glial cells and its predominant expression in hypoxic areas overexpressing HIF‑1α was highlighted. To investigate the role of ADNP on the HIF‑VEGF axis in GBM, a human U87MG GBM cell line was cultured with a hypoxic mimetic agent, deferoxamine, and cells were treated with the smallest active fragment of ADNP, known as NAP. The protein expression and distribution of HIF‑1α and VEGF was detected using western blot analysis and immunofluorescence assay. Results demonstrated that ADNP modulates the hypoxic‑angiogenic pathway in GBM cells by reducing VEGF secretion, detected through ELISA assay, as well as modulating their migration, assessed through wound healing assay. Although deeper investigation is necessary, the present study suggested that ADNP could be involved in PACAP anti‑invasive effects in GBM.
多形性胶质母细胞瘤（GBM）是一种影响成人、预后较差的脑癌。这是一种实体瘤，其特征是细胞迁移和侵袭率高。不受控制的细胞增殖在肿瘤块中形成缺氧微环境，这导致缺氧诱导因子（HIFs）的过度表达。这诱导血管内皮生长因子（VEGF）的激活，而VEGF负责不受控制的新生血管形成。最近的研究表明垂体腺苷酸环化酶激活肽（PACAP）在GBM中具有抗侵袭作用。PACAP对中枢神经系统的作用也通过活性依赖的神经保护蛋白（ADNP）激活来介导。迄今为止，尚无关于其在GBM中作用的证据。因此，对ADNP在GBM中的参与情况进行了研究。通过共聚焦显微镜分析人GBM样本中的ADNP表达，在大多数胶质细胞中检测到高ADNP免疫反应性，并突出显示其在过表达HIF-1α的缺氧区域中的主要表达。为了研究ADNP在GBM中对HIF-VEGF轴的作用，将人U87MG GBM细胞系用缺氧模拟剂去铁胺培养，并用ADNP的最小活性片段（称为NAP）处理细胞。使用蛋白质印迹分析和免疫荧光测定法检测HIF-1α和VEGF的蛋白质表达和分布。结果表明，ADNP通过减少ELISA检测到的VEGF分泌以及通过伤口愈合试验评估的细胞迁移来调节GBM细胞中的缺氧血管生成途径。尽管有必要进行更深入的研究，但本研究表明ADNP可能参与了PACAP在GBM中的抗侵袭作用。
