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A knock-in mouse model reveals roles for nuclear Apc in cell proliferation, Wnt signal inhibition and tumor suppression

Oncogene volume 31, pages 2423–2437 (2012)Cite this article

Abstract

Mutation of the tumor suppressor adenomatous polyposis coli (APC) is considered an initiating step in the genesis of the vast majority of colorectal cancers. APC inhibits the Wnt-signaling pathway by targeting the proto-oncogene β-catenin for destruction by cytoplasmic proteasomes. In the presence of a Wnt signal, or in the absence of functional APC, β-catenin can serve as a transcription cofactor for genes required for cell proliferation such as cyclin-D1 and c-Myc. In cultured cells, APC shuttles between the nucleus and the cytoplasm, with nuclear APC implicated in the inhibition of Wnt target gene expression. Adopting a genetic approach to evaluate the functions of nuclear APC in the context of a whole organism, we generated a mouse model with mutations that inactivate the nuclear localization signals (NLSs) of Apc (ApcmNLS). ApcmNLS/mNLS mice are viable and fractionation of mouse embryonic fibroblasts (MEFs) isolated from these mice revealed a significant reduction in nuclear Apc as compared with Apc+/+ MEFs. The levels of Apc and β-catenin protein were not significantly altered in small intestinal epithelia from ApcmNLS/mNLS mice. Compared with Apc+/+ mice, ApcmNLS/mNLS mice showed increased proliferation in epithelial cells from the jejunum, ileum and colon. These same tissues from ApcmNLS/mNLS mice showed more mRNA from three genes upregulated in response to canonical Wnt signal, c-Myc, axin-2 and cyclin-D1, and less mRNA from Hath-1, which is downregulated in response to Wnt. These observations suggest a role for nuclear Apc in the inhibition of canonical Wnt signaling and the control of epithelial proliferation in intestinal tissue. Furthermore, we found ApcMin/+ mice, which harbor a mutation that truncates Apc, to have an increased polyp size and multiplicity if they also carry the ApcmNLS allele. Taken together, this analysis of the novel ApcmNLS mouse model supports a role for nuclear Apc in the control of Wnt target genes, intestinal epithelial cell proliferation and polyp formation.

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Acknowledgements

This work was supported by RO1 CA10922 from the National Cancer Institute and 5P20 RR15563. We extend our gratitude to Alan Godwin, Kirk Thomas and Mario Capecchi for advice on the generation of the knock-in mouse and for the lambda phage library, and the tACE-Cre-Neor- and TkHSV-containing constructs. We thank Marc Roth, Areli Monarrez, Ashrita Abraham and Travis Friesen for technical assistance; the staff at the University of Kansas Animal Care Unit for excellent mouse husbandry and David Davido for critical reading of the manuscript. We also thank Reka Nagy and Drs Andras Nagy, Janet Rossant and Wanda Abramow-Newerly (Mount Sinai Hospital) for the mouse R1 ES cells.

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Authors and Affiliations

  1. Department of Molecular Biosciences, University of Kansas, Lawrence, KS, USA

    M Zeineldin, J Cunningham, W McGuinness, P Alltizer, B Blanchat & K L Neufeld

  2. Eccles Institute of Human Genetics, University of Utah, Salt Lake City, UT, USA

    B Cowley

  3. Department of Microbiology, University of Virginia, Charlottesville, VA, USA

    W Xu

  4. Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS, USA

    D Pinson

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  1. M Zeineldin
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Correspondence to K L Neufeld.

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Zeineldin, M., Cunningham, J., McGuinness, W. et al. A knock-in mouse model reveals roles for nuclear Apc in cell proliferation, Wnt signal inhibition and tumor suppression. Oncogene 31, 2423–2437 (2012). https://doi.org/10.1038/onc.2011.434

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