Abstract
Caspases mediate apoptosis and have also been implicated in stem-cell biology. How caspases are linked to stem-cell biology is not known. Here, we show that the apoptotic blebs of cancer cells fuse together to form novel structures called ‘blebbishields’. Blebbishields form spheres by fusion. Both blebbishield formation and sphere formation involve active caspases and N-linked glycosylation. Sphere formation is enhanced by acidic pH and is counteracted by inhibitors of proton pump, caspases, and cholesterol. The blebbishields from VEGFR2High cells are capable of enhanced sphere formation. Blebbishields express transiently downregulated stem-cell markers and the sphere-forming blebbishield-derived cells are tumorigenic. Our study demonstrates that the cancer stem cells can survive after apoptosis by blebbishield formation and subsequent sphere formation.
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Abbreviations
- p70S6K:
-
70-kDa ribosomal protein S6 kinase
- VEGF-A:
-
vascular endothelial growth factor-A
- VEGFR2:
-
vascular endothelial growth factor receptor-2
- GSK-3β:
-
glycogen synthase kinase-3β
- BSE-2:
-
blebbishield serial ejection-2
- CSCs:
-
cancer stem cells
- TSC-2:
-
tuberous sclerosis complex-2
- NEAA:
-
non-essential amino acids
- 4EBP1:
-
4E binding protein-1
- PARP:
-
poly-ADP ribose polymerase
- BE medium:
-
blebbishield ejection medium
- XIAP:
-
X-linked inhibitor of apoptosis
- PPI:
-
proton pump inhibitor
- ABCB1:
-
ATP-binding cassette subfamily B member-1
- CD44:
-
cluster of differentiation 44
- CK-20:
-
cytokeratin 20
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Acknowledgements
We sincerely thank Dr. David J McConkey for generously providing microarray data, cell lines, and for intellectual and editorial help with the manuscript, and Drs Bharat B. Aggarwal, Gomez-Manzano Candelaria, Felipe Samaniego, Isaiah J Fidler, Sun-Jin Kim, Guillermo Garcia-Manero, Xiongbin Lu, Chun Li, Gordon B Mills, and Molina Jennifer for providing cell lines/reagents, Dr. Robert R Langley and Mr. Kenneth Dunner Jr. for helping with electron microscopy, Mr. Kunal Mandal and Ms. Karen Ramirez for helping in cell sorting, Dr. Aron Mobley, Dr. Neema Navai and Ms. I-Ling Lee for various helps, Ms. Deming Stephanie for editorial assistance with the manuscript. GJG was supported by MD Anderson Cancer Center and the Clayton Foundation for Research. This research was supported in part by the National Institutes of Health through MD Anderson Cancer Center’s Support Grant, CA016672.
Author contributions
GJG designed, performed all experiments (except microarray), interpreted and documented results, and wrote the manuscript. WC contributed to the microarray experiment. JBS, EKL, DLW, and GJG performed the orthotopic mice experiments. AMK provided expert guidance in the design of experiments and interpretation of data, writing the manuscript, and generating the RT4v6 cell line.
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Jinesh, G., Choi, W., Shah, J. et al. Blebbishields, the emergency program for cancer stem cells: sphere formation and tumorigenesis after apoptosis. Cell Death Differ 20, 382–395 (2013). https://doi.org/10.1038/cdd.2012.140
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DOI: https://doi.org/10.1038/cdd.2012.140
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