Abstract
The mammalian homeobox transcription factor CDX2 has key roles in intestinal development and differentiation. Heterozygous Cdx2 mice develop one or two benign hamartomas in the proximal colon, whereas heterozygous ApcΔ716 mice develop numerous adenomatous polyps, mostly in the small intestine. Here we show that the colonic polyp number is about six times higher in Apc+/Δ716 Cdx2+/− compound mutant mice. Levels of both APC and CDX2 were significantly lower in the distal colon, which caused high anaphase bridge index (ABI) associated with a higher frequency of loss of heterozygosity (LOH) at Apc. In cultured rat intestinal epithelial and human colon cancer cell lines, suppression of CDX2 by antisense RNA caused marked increases in ABI and chromosomal aberrations. This was mediated by stimulation of the mTOR pathway, causing translational deregulation and G1-S acceleration, associated with low levels of p27 and activation of cyclin E–Cdk2. We obtained similar results in the colonic mucosa of Apc+/Δ716 Cdx2+/− compound mutant mice. Forced activation of mTOR through upstream regulator Akt also increased ABI in colon cancer cells. High ABI in all cell lines was suppressed by mTOR inhibitors LY294002 and rapamycin. These results suggest that reduced expression of CDX2 is important in colon tumorigenesis through mTOR-mediated chromosomal instability.
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Acknowledgements
We thank R. L. White and P. K. Vogt for valuable comments on the manuscript; F. Tamanoi for unpublished information; M. Sugai and A. Shimizu for help in flow cytometry; H. Seno and T. Ishikawa for antibody to mouse CDX2; J.-N. Freund for mouse Cdx2 cDNA; M. Tsujii for DLD-1 and IEC-6 cell lines; and M. Aoki, N. Harada, S. Yonehara, E. Nishida, T. Ishikawa and H. Miyoshi for help, suggestions and discussions. This work was supported by grants from Organization for Pharmaceutical Safety and Research and the Ministry of Education, Culture, Sports, Science, and Technology, Japan.
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Aoki, K., Tamai, Y., Horiike, S. et al. Colonic polyposis caused by mTOR-mediated chromosomal instability in Apc+/Δ716 Cdx2+/− compound mutant mice. Nat Genet 35, 323–330 (2003). https://doi.org/10.1038/ng1265
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DOI: https://doi.org/10.1038/ng1265
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