Three-dimensional (3D) tumor spheroid invasion assay.
Three-dimensional (3D) tumor spheroid invasion assay.
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DOI：
10.3791/52686
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发表时间：
2015-05-01
期刊：
影响因子：
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通讯作者：
Eccles SA
中科院分区：
文献类型：
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作者：
Vinci M;Box C;Eccles SA
周围正常组织的侵袭通常被认为是恶性（与良性相反）肿瘤的关键标志。特别是对于某些癌症（例如，脑肿瘤，如多形性胶质母细胞瘤和头颈部鳞状细胞癌（SCCHN），它是严重发病的原因，即使没有远处转移，也可能危及生命。此外，不幸的是，治疗后复发的癌症通常表现为更具侵袭性的表型。因此，有机会靶向侵袭过程，以提供可以补充标准抗增殖剂的新疗法。到目前为止，这种策略一直受到缺乏强大的，可重复的检测方法，适合详细分析入侵和药物筛选的阻碍。在这里，我们提供了一个简单的微板方法（基于均匀的，自组装的3D肿瘤球体），这类研究具有很大的潜力。我们使用人胶质母细胞瘤细胞系和SCCHN模型举例说明了该检测平台，其中对靶向表皮生长因子受体（EGFR）抑制剂的耐药性的发展与增强的基质侵袭潜力相关。我们还提供了两种半自动定量的替代方法：一种使用成像细胞仪，第二种只需要标准的显微镜和数字图像分析的图像捕获。
Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in particular (e.g., brain tumors such as glioblastoma multiforme and squamous cell carcinoma of the head and neck – SCCHN) it is a cause of severe morbidity and can be life-threatening even in the absence of distant metastases. In addition, cancers which have relapsed following treatment unfortunately often present with a more aggressive phenotype. Therefore, there is an opportunity to target the process of invasion to provide novel therapies that could be complementary to standard anti-proliferative agents. Until now, this strategy has been hampered by the lack of robust, reproducible assays suitable for a detailed analysis of invasion and for drug screening. Here we provide a simple micro-plate method (based on uniform, self-assembling 3D tumor spheroids) which has great potential for such studies. We exemplify the assay platform using a human glioblastoma cell line and also an SCCHN model where the development of resistance against targeted epidermal growth factor receptor (EGFR) inhibitors is associated with enhanced matrix-invasive potential. We also provide two alternative methods of semi-automated quantification: one using an imaging cytometer and a second which simply requires standard microscopy and image capture with digital image analysis.