Quantitative determination of apoptotic death in cultured human pancreatic cancer cells by propidium iodide and digitonin
Quantitative determination of apoptotic death in cultured human pancreatic cancer cells by propidium iodide and digitonin
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DOI：
10.1016/s0304-3835(99)00107-x
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发表时间：
1999-08-03
期刊：
影响因子：
9.7
通讯作者：
Tanaka, M
中科院分区：
文献类型：
--
作者：
Zhang, L;Mizumoto, K;Tanaka, M
我们在这里介绍了一种体外细胞毒性试验的有效性，它可以快速定量地测量死亡细胞的凋亡数和细胞的生长率。用多孔板阅读器测定了依托泊苷作用下培养的人胰腺癌细胞经洋地黄素处理后，碘化丙啶(PI)对死亡细胞和总细胞的荧光强度。该方法测定的死亡细胞百分率与台盼蓝染色测定的细胞死亡百分率有很好的相关性。此外，同时测定的细胞生长率也与显微镜直接计数的细胞数量相关。我们证明这种最初用于评估坏死的方法可以应用于测量细胞的凋亡性死亡。综上所述，这种简单的检测方法对于测试抗癌药物的疗效和研究培养细胞中细胞凋亡的分子机制是有用的。(C)1999爱思唯尔爱尔兰科学有限公司。保留所有权利。
We present here the efficacy of an in vitro cytotoxicity assay which can measure rapidly both apoptotic dead cells and cell growth rate, quantitatively. Using a multi-well plate reader, the fluorescence intensity of propidium iodide (PI) corresponding to dead cells and to total cells after digitonin treatment were measured in cultured human pancreatic cancer cells following exposure to etoposide. The percentage of dead cells measured by this assay was well correlated to that determined by Trypan blue staining. Furthermore, the cell growth rate determined simultaneously was also correlated to the cell number counted directly using a microscope. We demonstrate that this method, which was originally established for evaluating necrosis, could be applied to measure apoptotic cell death. Taken together, this simple assay is useful for testing the efficacy of anti-cancer agents and for investigating the molecular mechanisms of apoptosis in the cultured cells. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.