PO17A NOVEL IMMUNOHISTOCHEMICAL METHODOLOGY FOR THE INVESTIGATION OF STEM CELL-LIKE SUBPOPULATIONS IN GLIOMA SPHEROIDS AND MIGRATORY CELLS
PO17A NOVEL IMMUNOHISTOCHEMICAL METHODOLOGY FOR THE INVESTIGATION OF STEM CELL-LIKE SUBPOPULATIONS IN GLIOMA SPHEROIDS AND MIGRATORY CELLS
复制标题
PO17A 用于研究胶质瘤球体和迁移细胞中干细胞样亚群的新型免疫组织化学方法
DOI：
10.1093/neuonc/nov284.14
复制
发表时间：
2015
期刊：
影响因子：
15.9
通讯作者：
Cheng V
中科院分区：
文献类型：
--
作者：
Cheng V
引言在高度浸润性肿瘤（如胶质母细胞瘤）的标准治疗中，靶向肿瘤侵袭可能是一种补充治疗。侵袭性被认为与胶质瘤细胞的不同亚群的干细胞样品质有关。侵袭性和非侵袭性细胞之间表型差异的表征将有助于特异性抗迁移药物的开发。一种新的免疫组化方法被用来在3D球体侵袭assay. METHODS从U87和U251细胞产生的球体包埋在胶原基质中，并与抗迁移药物氯化锂和生物靛玉红处理72小时后，表型的侵袭性迁移胶质瘤细胞治疗后。建立了制备胶原包埋的球状体和迁移细胞用于免疫组织化学（IHC）的方案。在U87和U251中，保持在原始核心中的细胞中的蛋白质表达谱与迁移细胞中的蛋白质表达谱不同。在U251中观察到SOX-2表达的最显著变化，其中药物处理导致核心维持细胞中SOX-2水平降低，而迁移细胞表现出更高水平的SOX-2。我们也可以量化药物诱导的变化，从这两个细胞系产生的球状体中的增殖和凋亡事件，使用这种method.CONCLUSIONA新的IHC协议允许在3D实验模型中的细胞的侵袭行为的调查。干性标志物的蛋白质表达谱表明，胶质瘤球体内存在细胞亚群，这些细胞亚群对抗迁移药物的反应可能不同。
INTRODUCTIONTargeting tumour invasion may represent a complementary treatment when added to standard care in highly infiltrating tumours such as glioblastomas. Invasiveness is believed to be related to stem cell-like qualities of distinct subpopulations of glioma cells. The characterisation of phenotypic differences between invading and noninvading cells will aid the development of specific anti-migratory drugs. A novel immunohistochemical approach was taken to phenotypically characterise invasive migratory glioma cells after drug treatment in a 3D spheroid invasion assay.METHODSpheroids generated from U87 and U251 cells were embedded in a collagen matrix and treated over 72 hours with the antimigratory drugs LiCl and Bio-Indirubin. A protocol was established to prepare the collagen embedded spheroids and migratory cells for immunohistochemistry (IHC). Markers of proliferation, apoptosis and stemness were optimised for IHC.RESULTSIn both U87 and U251, protein expression profiles in cells maintained in the original core were distinct from profiles in migratory cells. The most striking change of expression was observed for SOX-2 in U251, where drug treatment led to a reduction of SOX-2 levels in core-maintained cells whereas migrating cells exhibited higher levels of SOX-2. We could also quantify drug-induced changes in proliferation and apoptotic events in spheroids generated from both cell lines using this method.CONCLUSIONA novel IHC protocol allowed the investigation of the invasive behaviour of cells in a 3D experimental model. Protein expression profiles of markers of stemness indicate that subpopulations of cells exist within glioma spheroids that may respond differently to anti-migratory drugs.