Department of Molecular Biotechnology and Microbiology, University of Debrecen, Debrecen, Hungary.
Department of Pharmaceutical Chemistry, University of Debrecen, Debrecen, Hungary.
Anticancer Res. 2021 Jan;41(1):137-149. doi: 10.21873/anticanres.14759.
BACKGROUND/AIM: Conventional viability tests, help to screen the cellular effects of candidate molecules, but the endpoint of these measurements lacks sufficient information regarding the molecular aspects. A non-invasive, easy-to-setup live-cell microscopic method served to in-depth analysis of mechanisms of potential anticancer drugs.
The proposed method combining the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test with time-lapse scanning microscopy (TLS), provided additional data related to the cell-cycle and the dynamic properties of cell morphology. Apoptotic and necrotic events became detectable with these methods.
Quantification of the results was assisted by image analysis of the acquired image sequences. After demonstrating the potential of the TLS method, a series of experiments compared the in vitro effect of a known and a newly synthesized nucleoside analogue.
The proposed approach provided a more in-depth insight into the cellular processes that can be affected by known chemotherapeutic agents including nucleoside analogues rather than applying repeated individual treatments.
背景/目的：传统的生存能力测试有助于筛选候选分子的细胞效应，但这些测量的终点缺乏关于分子方面的足够信息。一种非侵入性、易于设置的活细胞显微镜方法可用于深入分析潜在抗癌药物的作用机制。
该方法将 MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）测试与时间 lapse 扫描显微镜（TLS）相结合，提供了与细胞周期和细胞形态动力学特性相关的附加数据。通过这些方法可以检测到细胞凋亡和坏死事件。
通过对获取的图像序列进行图像分析，辅助对结果进行定量。在证明了 TLS 方法的潜力之后，进行了一系列实验比较了一种已知和一种新合成的核苷类似物的体外效应。
与应用重复的单独处理相比，所提出的方法可以更深入地了解受包括核苷类似物在内的已知化疗药物影响的细胞过程。
