Skvortsov D A, Kalinina M A, Zhirkina I V, Vasilyeva L A, Ivanenkov Y A, Sergiev P V, Dontsova O A
Chemistry Department, Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, Russia.
Faculty of Biology and Biotechnologies, Higher School of Economics, Moscow, Russia.
Front Pharmacol. 2021 Oct 11;12:713103. doi: 10.3389/fphar.2021.713103. eCollection 2021.
For the search of anticancer compounds in modern large chemical libraries, new approaches are of great importance. Cocultivation of the cells of tumor and non-tumor etiology may reveal specific action of chemicals on cancer cells and also take into account some effects of the tumor cell's microenvironment. The fluorescent cell cocultivation test (FCCT) has been developed for screening of substances that are selectively cytotoxic on cancerous cells. It is based on the mixed culture of lung carcinoma cells A549'_EGFP and noncancerous fibroblasts of lung VA13_Kat, expressing different fluorescent proteins. Analysis of the cells was performed with the high-resolution scanner to increase the detection rate. The combination of cocultivation of cells with scanning of fluorescence reduces the experimental protocol to three steps: cells seeding, addition of the substance, and signal detection. The FCCT analysis does not disturb the cells and is compatible with other cell-targeted assays. The suggested method has been adapted for a high-throughput format and applied for screening of 2,491 compounds. Three compounds were revealed to be reproducibly selective in the FCCT although they were invisible in cytotoxicity tests in individual lines. Six structurally diverse indole, coumarin, sulfonylthiazol, and rifampicin derivatives were found and confirmed with an independent assay (MTT) to be selectively cytotoxic to cancer cells in the studied model.
在现代大型化学文库中寻找抗癌化合物时，新方法至关重要。肿瘤和非肿瘤病因细胞的共培养可能揭示化学物质对癌细胞的特定作用，同时也能考虑到肿瘤细胞微环境的一些影响。荧光细胞共培养试验（FCCT）已被开发用于筛选对癌细胞具有选择性细胞毒性的物质。它基于表达不同荧光蛋白的肺癌细胞A549'_EGFP和肺非癌成纤维细胞VA13_Kat的混合培养。使用高分辨率扫描仪对细胞进行分析以提高检测率。细胞共培养与荧光扫描相结合将实验方案简化为三个步骤：细胞接种、添加物质和信号检测。FCCT分析不会干扰细胞，并且与其他细胞靶向检测兼容。所建议的方法已适用于高通量形式，并应用于2491种化合物的筛选。尽管在单个细胞系的细胞毒性试验中未发现这三种化合物，但在FCCT中发现它们具有可重复的选择性。发现了六种结构多样的吲哚、香豆素、磺酰噻唑和利福平衍生物，并通过独立试验（MTT）证实它们在所研究的模型中对癌细胞具有选择性细胞毒性。
