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Different phenotypic consequences of simultaneous versus stepwise Apc loss

Oncogene volume 31, pages 2028–2038 (2012)Cite this article

Abstract

APC is considered a gatekeeper for colorectal cancer (CRC). Cells with heterozygous APC mutations have altered expression profiles suggesting that the first APC hit may help set the stage for subsequent transformation. Therefore, we measured transformation efficiency following what we have designated as ‘simultaneous’ versus ‘stepwise’ Apc loss. We combined a conditional Apc allele (ApcCKO) with a Cre reporter gene and an out-of-frame Cre allele (Pms2cre) that stochastically becomes functional by a frameshift mutation in single cells. Loss of one Apc allele (ApcCKO/+) had little consequence, whereas simultaneous loss of both Apc alleles (ApcCKO/CKO) resulted in increased clonal expansion (crypt fission), consistent with the gatekeeper function of Apc. Interestingly, our analyses showed that most of the Apc-deficient crypts in ApcCKO/CKO mice appeared normal, with morphological transformation, including β-catenin deregulation, occurring in only 17% of such crypts. To determine whether transformation efficiency was different following stepwise Apc loss, we combined ApcCKO with a germline mutant allele, either ApcMin or Apc1638N. Transformation efficiency following stepwise Apc loss (ApcMin/CKO or Apc1638N/CKO) was increased five-fold and essentially all of the Apc-deficient cells were dysplastic. In summary, our data suggest that the gatekeeper function of Apc consists of two roles, clonal expansion and morphological transformation, because simultaneous Apc loss frequently leads to occult clonal expansion without morphological transformation, whereas stepwise Apc loss more often results in visible neoplasia. Finally, that Apc-deficient cells in certain scenarios can retain a normal phenotype is unexpected and may have clinical implications for surveillance strategies to prevent CRC.

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Acknowledgements

We thank Drs James Stringer and Melissa Wong for critical reading of the manuscript. We also thank Drs Raju Kucherlapati and Winfried Edelmann for the ApcCKO and Apc1638N mice, respectively; John Swain and Dr Melissa Wong for ApcMin mice and Apc580S intestinal material; and Dan Lioy and Drs Gail Mandel and Paul Brehm for assistance with confocal microscopy. RML and DS were funded by NIH Grant 2R01GM032741-28. JMF was funded by NIH training Grant 5T32HD046420-05 and ACS postdoctoral fellow PF-11-067-01-TBE.

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  1. Molecular and Medical Genetics, Oregon Health and Science University, Portland, OR, USA

    J M Fischer, A J Miller & R M Liskay

  2. Department of Pathology, Norris Cancer Center, University of Southern California School of Medicine, Los Angeles, CA, USA

    D Shibata

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  1. J M Fischer
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  2. A J Miller
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Correspondence to R M Liskay.

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Fischer, J., Miller, A., Shibata, D. et al. Different phenotypic consequences of simultaneous versus stepwise Apc loss. Oncogene 31, 2028–2038 (2012). https://doi.org/10.1038/onc.2011.385

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