Research Institute of Pharmaceutical Sciences and College of Pharmacy, Seoul National University, Seoul, 151-742, South Korea.
Lab Chip. 2011 Jan 7;11(1):104-14. doi: 10.1039/c0lc00110d. Epub 2010 Nov 8.
Drug-induced cardiotoxicity or cytotoxicity followed by cell death in cardiac muscle is one of the major concerns in drug development. Herein, we report a high-content quantitative multicolor single cell imaging tool for automatic screening of drug-induced cardiotoxicity in an intact cell. A tunable multicolor imaging system coupled with a miniaturized sample platform was destined to elucidate drug-induced cardiotoxicity via simultaneous quantitative monitoring of intracellular sodium ion concentration, potassium ion channel permeability and apoptosis/necrosis in H9c2(2-1) cell line. Cells were treated with cisapride (a human ether-à-go-go-related gene (hERG) channel blocker), digoxin (Na(+)/K(+)-pump blocker), camptothecin (anticancer agent) and a newly synthesized anti-cancer drug candidate (SH-03). Decrease in potassium channel permeability in cisapride-treated cells indicated that it can also inhibit the trafficking of the hERG channel. Digoxin treatment resulted in an increase of intracellular [Na(+)]. However, it did not affect potassium channel permeability. Camptothecin and SH-03 did not show any cytotoxic effect at normal use (≤300 nM and 10 μM, respectively). This result clearly indicates the potential of SH-03 as a new anticancer drug candidate. The developed method was also used to correlate the cell death pathway with alterations in intracellular [Na(+)]. The developed protocol can directly depict and quantitate targeted cellular responses, subsequently enabling an automated, easy to operate tool that is applicable to drug-induced cytotoxicity monitoring with special reference to next generation drug discovery screening. This multicolor imaging based system has great potential as a complementary system to the conventional patch clamp technique and flow cytometric measurement for the screening of drug cardiotoxicity.
药物诱导的心肌细胞毒性或细胞毒性导致心肌细胞死亡是药物开发中的主要关注点之一。在此，我们报告了一种高通量定量多色单细胞成像工具，用于自动筛选完整细胞中的药物诱导的心脏毒性。一个可调多色成像系统与一个微型化的样品平台相结合，旨在通过同时定量监测细胞内钠离子浓度、钾离子通道通透性和 H9c2(2-1)细胞系中的细胞凋亡/坏死，来阐明药物诱导的心脏毒性。用 cisapride（一种人 ether-à-go-go 相关基因 (hERG) 通道阻滞剂）、地高辛（Na(+)/K(+)-pump 阻滞剂）、喜树碱（抗癌剂）和一种新合成的抗癌药物候选物（SH-03）处理细胞。 cisapride 处理的细胞中钾离子通道通透性降低表明它也可以抑制 hERG 通道的运输。地高辛处理导致细胞内 [Na(+)] 增加。然而，它并没有影响钾离子通道通透性。喜树碱和 SH-03 在正常使用时（分别为≤300 nM 和 10 μM）均没有显示出任何细胞毒性作用。这一结果清楚地表明了 SH-03 作为一种新的抗癌药物候选物的潜力。所开发的方法还用于将细胞死亡途径与细胞内 [Na(+)] 的变化相关联。所开发的方案可以直接描绘和定量靶向细胞反应，随后提供一种自动化、易于操作的工具，适用于药物诱导的细胞毒性监测，特别是与下一代药物发现筛选相关。这种基于多色成像的系统具有成为传统膜片钳技术和流式细胞术测量的补充系统的巨大潜力，可用于药物心脏毒性的筛选。
