High-content analysis of tumour cell invasion in three-dimensional spheroid assays.
High-content analysis of tumour cell invasion in three-dimensional spheroid assays.
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DOI：
10.18632/oncoscience.171
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Brüning-Richardson A
中科院分区：
文献类型：
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作者：
Cheng V;Esteves F;Chakrabarty A;Cockle J;Short S;Brüning-Richardson A
靶向浸润性肿瘤细胞是对抗癌症侵袭和转移的有吸引力的方式。在这里，我们描述了一种新的和可重复的方法，用于高含量的入侵细胞的分析，在一个高级别的胶质瘤模型中使用多细胞肿瘤球体测定。从胶质瘤细胞系U87和U251产生的球状体的活细胞成像，提供了对响应于抗迁移药物的迁移动力学和细胞形态的洞察。免疫荧光成像证实了细胞骨架重排在处理的细胞，表明对细胞形态的直接影响。通过来自明场图像的迁移指数（MI）确定对迁移的影响，该图像证实了药物的抗迁移活性。还观察到了对核心的显著影响，提示了无序增殖。一种新开发的制备球状体和迁移细胞用于免疫组织化学的技术允许用选择的标志物评估对药物治疗的反应。在核心内维持的细胞和迁移细胞之间注意到蛋白质表达的差异，这表明球状体核心内存在细胞亚群。我们的结论是，这种高含量的分析使研究人员能够进行抗肿瘤侵袭化合物的筛选，并首次研究它们对细胞动力学的影响，特别是与蛋白质表达有关的影响。
Targeting infiltrating tumour cells is an attractive way of combating cancer invasion and metastasis. Here we describe a novel and reproducible method for high content analysis of invading cells using multicellular tumour spheroid assays in a high grade glioma model. Live cell imaging of spheroids generated from glioma cell lines, U87 and U251, gave insight into migration dynamics and cell morphology in response to anti-migratory drugs. Immunofluorescence imaging confirmed cytoskeletal rearrangements in the treated cells indicating a direct effect on cell morphology. Effect on migration was determined by a Migration Index (MI) from brightfield images which confirmed anti-migratory activity of the drugs. A marked effect on the core with treatment suggestive of disordered proliferation was also observed. A newly developed technique to prepare the spheroids and migratory cells for immunohistochemistry allowed an assessment of response to drug treatment with a selection of markers. A difference in protein expression was noted between cells maintained within the core and migratory cells indicative of the presence of cell subpopulations within the spheroid core. We conclude that this high content analysis allows researchers to perform screening of anti-tumour invasion compounds and study their effects on cellular dynamics, particularly in relation to protein expression, for the first time.