Relationship between intracellular pH, metabolic co-factors and caspase-3 activation in cancer cells during apoptosis
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 o...
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Published in | Biochimica et biophysica acta. Molecular cell research Vol. 1864; no. 3; pp. 604 - 611 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Netherlands
Elsevier B.V
01.03.2017
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Subjects | |
Online Access | Get full text |
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Summary: | 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.
•Multiparameter fluorescence microscopy is applied for apoptosis study in cancer cells.•Genetically encoded sensors are used for pHi and caspase-3 activation imaging.•Metabolic co-factors, pHi and caspase-3 activity are visualized in apoptotic cells.•Dynamic changes in metabolism, pHi and caspase-3 activity are shown in apoptosis. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0167-4889 1879-2596 |
DOI: | 10.1016/j.bbamcr.2016.12.022 |