Abstract
The transcription factor signal transducer and activator of transcription 3 (STAT3) is frequently activated in human cancers. Interestingly, STAT3 also maintains the pluripotency and self-renewal of murine embryonic stem cells, and several tissue stem cell types. To investigate whether STAT3 also maintains the small-intestine crypt stem cell, we conditionally inactivated a Floxed Stat3 allele (Stat3fl) in murine small-intestine crypt stem cells. Following Cre recombinase expression, apoptosis increased in Stat3fl/− experimental crypts relative to Stat3wt/− controls before declining. Control Stat3wt/− mice carrying a Flox-STOP LacZ reporter transgene stably expressed LacZ after Cre induction. In contrast, Stat3fl/− intestine LacZ expression initially increased modestly, before declining to background levels. Quantitative PCRs revealed a similar transient in recombined Stat3fl allele levels. Long-term bromodeoxyuridine labelling directly demonstrated that functional STAT3 is required for +4 to +6 region label-retaining small-intestine stem cell survival. Rapid clearance of recombined Stat3fl/− cells involves apoptosis potentially induced by elevated c-Myc in non-recombined cells and involves elevated p53 expression and caspase 3 activation. Intriguingly, Stat3fl/− intestine recombination triggered dramatically upregulated polycomb transcriptional repressor Bmi1 – potentially accelerating recombined crypt repopulation. In summary, STAT3 activity is absolutely required for small-intestine crypt stem cell survival at both the +4 to +6 label-retaining and crypt base columnar cell locations.
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Abbreviations
- AH-Cre:
-
Cyp1A1 promoter-Cre recombinase
- BrdU:
-
bromodeoxyuridine
- CBC:
-
crypt base columnar cell
- Ct:
-
threshold cycle
- DAPI:
-
4′,6-diamidino-2-phenylindole
- Fl:
-
floxed
- H+E:
-
haematoxylin plus eosin
- HRP:
-
horseradish peroxidase
- mES:
-
murine embryonic stem cell
- NGS:
-
normal goat serum
- PcG:
-
polycomb group
- Q-PCR:
-
quantitative polymerase chain reaction
- SH2:
-
src homology 2
- STAT:
-
signal transducer and activator of transcription
- TBS:
-
Tris-buffered saline
- WT:
-
wild type
- –:
-
null
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Acknowledgements
This work was supported by a Cancer Research UK programme grant, we also gratefully thank M Bishop for providing mouse genotyping services, A Hayes for performing confocal fluorescence immunohistochemistry, C Oliver for preliminary data and thank L Parry for providing two cycles of Cre induction small-intestine slides.
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Matthews, J., Sansom, O. & Clarke, A. Absolute requirement for STAT3 function in small-intestine crypt stem cell survival. Cell Death Differ 18, 1934–1943 (2011). https://doi.org/10.1038/cdd.2011.77
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DOI: https://doi.org/10.1038/cdd.2011.77
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