Relationship between intracellular pH, metabolic co-factors and caspase-3 activation in cancer cells during apoptosis
Relationship between intracellular pH, metabolic co-factors and caspase-3 activation in cancer cells during apoptosis
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DOI：
10.1016/j.bbamcr.2016.12.022
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发表时间：
2017-03-01
影响因子：
5.1
通讯作者：
Zagaynova, Elena V.
中科院分区：
文献类型：
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作者：
Sergeeva, Tatiana F.;Shirmanova, Marina V.;Zagaynova, Elena V.
凋亡细胞中发生复杂的分子事件级联，但细胞间的变异性使不同过程之间的顺序和互连的确定变得非常复杂。为了更好地理解程序性细胞死亡的机制，需要动态同时记录多个参数。在本文中，我们使用多参数荧光显微镜分析了星形孢菌素诱导细胞凋亡过程中体外活癌细胞的能量代谢、细胞内 pH 和 caspase-3 激活。我们对两种辅助因子 NAD(P)H 和 FAD 进行了代谢成像，并使用基因编码的 pH 指示剂 SypHer1 和基于 FRET 的 caspase-3 活性传感器 mKate2-DEVD-iRFP，通过共焦荧光显微镜和双光子荧光寿命成像显微镜可视化这些参数。研究CT26癌细胞凋亡过程中能量代谢、细胞内pH值和caspase-3激活之间的相关性及其动态变化。细胞凋亡的诱导伴随着氧化磷酸化、胞质酸化和 caspase-3 激活的转变。我们发现细胞质 pH 值的改变和氧化磷酸化的激活是与诱导细胞凋亡相关的相对早期的事件。 (C) 2017 Elsevier B.V. 保留所有权利。
A complex cascade of molecular events occurs in apoptotic cells but cell-to-cell variability significantly complicates determination of the order and interconnections between different processes. For better understanding of the mechanisms of programmed cell death, dynamic simultaneous registration of several parameters is required. In this paper we used multiparameter fluorescence microscopy to analyze energy metabolism, intracellular pH and caspase-3 activation in living cancer cells in vitro during staurosporine-induced apoptosis. We performed metabolic imaging of two co-factors, NAD(P)H and FAD, and used the genetically encoded pH-indicator SypHer1 and the FRET-based sensor for caspase-3 activity, mKate2-DEVD-iRFP, to visualize these parameters by confocal fluorescence microscopy and two-photon fluorescence lifetime imaging microscopy. The correlation between energy metabolism, intracellular pH and caspase-3 activation and their dynamic changes were studied in CT26 cancer cells during apoptosis. Induction of apoptosis was accompanied by a switch to oxidative phosphorylation, cytosol acidification and caspase-3 activation. We showed that alterations in cytosolic pH and the activation of oxidative phosphorylation are relatively early events associated with the induction of apoptosis. (C) 2017 Elsevier B.V. All rights reserved.