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Colonic polyposis caused by mTOR-mediated chromosomal instability in Apc+/Δ716 Cdx2+/− compound mutant mice

Nature Genetics volume 35, pages 323–330 (2003)Cite this article

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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Figure 1: Colonic polyps in Apc+/Δ716 Cdx2+/− mice.
Figure 2: Colonic expression of Cdx2 and Apc in ApcΔ716, Apc+/Δ716 Cdx2+/− and Cdx2+/− mice.
Figure 3: Anaphase cell analysis in ApcΔ716, Apc+/Δ716 Cdx2+/− and Cdx2+/− mice.
Figure 4: Anaphase bridges in cultured cells induced by suppression of CDX2 expression.
Figure 5: G1-S progression in low CDX2 clones.
Figure 6: Protein analyses in CDX2 DLD-1 clones and colonic mucosa and colonic cell kinetics in vivo.
Figure 7: Effects of kinase inhibitors on ABI in low CDX2 DLD-1 cells.

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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We thank R. L. White and P. K. Vogt for valuable comments on the manuscript; F. Tamanoi for unpublished information; M. Sugai and A. Shimizu for help in flow cytometry; H. Seno and T. Ishikawa for antibody to mouse CDX2; J.-N. Freund for mouse Cdx2 cDNA; M.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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Authors and Affiliations

  1. Department of Pharmacology, Graduate School of Medicine, Kyoto University, Yoshida-Konoé-cho, Sakyo-ku, Kyoto, 606-8501, Japan

    Koji Aoki, Masanobu Oshima & Makoto M Taketo

  2. Banyu Tsukuba Research Institute (Merck), Ibaraki, 300-2611, Japan

    Yoshitaka Tamai

  3. Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan

    Shigeo Horiike

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Correspondence to Makoto M Taketo.

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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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