Abstract
The BCL2 family of proteins regulate apoptosis by controlling mitochondrial outer membrane permeability. However, the effects on mitochondrial structure and bioenergetics have also been reported. Here we comprehensively characterized the effects of BCL2 and BCL(X)L on cellular energetics in MCF7 breast cancer cells using time-lapse confocal single-cell imaging and mitochondrial and cytosolic FRET reporters. We found that BCL2 and BCL(X)L increase the metabolic robustness of MCF7 cells, and that this was associated with increased mitochondrial NAD(P)H and ATP levels. Experiments with the F1F0 synthase inhibitor oligomycin demonstrated that BCL2 and in particular BCL(X)L, while not affecting ATP synthase activity, more efficiently coupled the mitochondrial proton motive force with ATP production. This metabolic advantage was associated with an increased resistance to nutrient deprivation and enhanced clonogenic survival in response to metabolic stress, in the absence of profound effects on cell death. Our data suggest that a primary function of BCL(X)L and BCL2 overexpression in tumor cells is to increase their resistance to metabolic stress in the tumor microenvironment, independent of cell death signaling.
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Data availability
The raw and processed RNA sequencing data for the MCF7 cell lines generated in this study are publicly available in GEO (reference number GSE158808).
Code availability
Processing and analysis code for the transcriptomic-based analysis is publicly available and archived at Zenodo (https://doi.org/10.5281/zenodo.4058036).
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Acknowledgements
We thank Dr Claus Reimertz for generating the BCL2- and BCL(X)L-overexpressing clones, and Dr Bram Boeckx for RNA sequencing. This research was funded by grants from the Irish Cancer Society Collaborative Cancer Research Centre BREAST-PREDICT (CCRC13GAL) and Science Foundation Ireland and the Health Research Board (13/IA/1881, 16/US/3301) to JHMP. We kindly thank Prof Hiroyuki Noji (Osaka University) and Prof. Hiromi Imamura (Kyoto University) for providing the mitoATeam and cytosolic ATeam plasmids. We also thank Luise Halang and Aisling O’Brien for technical assistance.
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Conception and design: FL, HD, and JHMP; Acquisition of data: FL, HD, MS, AUL, and DL; Writing, review and/or revision of the paper: FL, MS, HD, and JHMP; Study supervision: JHMP
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Lucantoni, F., Salvucci, M., Düssmann, H. et al. BCL(X)L and BCL2 increase the metabolic fitness of breast cancer cells: a single-cell imaging study. Cell Death Differ 28, 1512–1531 (2021). https://doi.org/10.1038/s41418-020-00683-x
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DOI: https://doi.org/10.1038/s41418-020-00683-x
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