Department of Oncology-Pathology, Karolinska Institute, S-17176 Stockholm, Sweden.
Chem Biol Drug Des. 2011 Oct;78(4):547-57. doi: 10.1111/j.1747-0285.2011.01170.x. Epub 2011 Jul 29.
Cell-based anticancer drug screening generally utilizes rapidly proliferating tumour cells grown as monolayer cultures. Hit compounds from such screens are not necessarily effective on hypoxic and slowly proliferating cells in 3-D tumour tissue. The aim of this study was to examine the potential usefulness of 3-D cultured tumour cells for anticancer drug screening. We used colon carcinoma multicellular spheroids containing hypoxic and quiescent cells in core areas for this purpose. Three libraries (∼11 000 compounds) were screened using antiproliferative activity and/or apoptosis as end-points. Screening of monolayer and spheroid cultures was found to identify different sets of hit compounds. Spheroid screening enriched for hydrophobic compounds: median XLogP values of 4.3 and 4.4 were observed for the hits in two independent screening campaigns. Mechanistic analysis revealed that the majority of spheroid screening hits were microtubuli inhibitors. One of these inhibitors was examined in detail and found to be effective against non-dividing cells in the hypoxic centres of spheroids. Spheroid screening represents a conceptually new strategy for anticancer drug discovery. Our findings have implications for drug library design and hit selection in projects aimed to develop drugs for the treatment of solid tumours.
基于细胞的抗癌药物筛选通常利用作为单层培养物生长的快速增殖肿瘤细胞。来自此类筛选的命中化合物在 3-D 肿瘤组织中的缺氧和缓慢增殖细胞上不一定有效。本研究旨在研究 3-D 培养的肿瘤细胞在抗癌药物筛选中的潜在用途。为此，我们使用含有核心区域缺氧和静止细胞的结肠癌细胞多细胞球体。使用增殖活性和/或细胞凋亡作为终点，对三个文库（约 11000 种化合物）进行了筛选。发现单层和球体培养物的筛选可识别出不同的命中化合物集。球体筛选富集了疏水性化合物：在两个独立的筛选实验中，命中化合物的中位 XLogP 值分别为 4.3 和 4.4。机制分析表明，大多数球体筛选命中化合物是微管抑制剂。其中一种抑制剂进行了详细研究，发现它对球体缺氧中心的非分裂细胞有效。球体筛选代表了抗癌药物发现的一种全新策略。我们的发现对旨在开发治疗实体瘤药物的项目中的药物库设计和命中化合物选择具有影响。
