Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Epithelial proliferation induces novel changes in APC expression

Oncogene volume 24, pages 6709–6718 (2005)Cite this article

Abstract

The role of wild-type adenomatous polyposis coli (APC) protein in native epithelia is poorly understood. The present study examined the relationships between wild-type APC and β-catenin expression in an established model of hyperproliferation, transmissible murine colonic hyperplasia (TMCH). Distal colonic crypts isolated from normal or TMCH mice were: (i) fractionated into cytosolic and nuclear components for Western blotting and immunoprecipitation (IP), (ii) extracted for total RNA isolation for Northern blotting and, (iii) analysed immunohistochemically by confocal microscopy. Western blots performed sequentially through day 12 TMCH with N-terminal APC antibodies revealed increased abundance of ∼312 kDa (p312) protein by day 6 (4.0±0.75-fold, n=6) that peaked by day 9, before declining by day 12. A ∼130 kDa (p130) band appeared at day 9 and increased by day 12 (1.5±0.11-fold, n=6). A C-terminal antibody detected only p312. APC mRNA level did not change during TMCH and appearance of p130 was not due to alternative splicing. Co-IP with N-terminal anti-APC antibodies, revealed APC's association with β-catenin both at day 6 and day 12. p130, but not p312, associated predominantly with β-catenin at day 12 during co-IP with anti-β-catenin. p130 also selectively accumulated in the nucleus, bound to nuclear β-catenin at day 12. Immunocytochemistry with N-terminal antibodies revealed an increasing crypt base : surface gradient of APC within the apical pole/apical-lateral membranes at day 6. At day 12, intense apical/cytoplasmic and occasional nuclear staining along the longitudinal crypt axis was observed. Full-length APC increases during epithelial hyperproliferation and may represent a homoeostatic response. The dramatic increase in cytoplasmic and sporadic nuclear APC staining at day 12 with N-terminal antibodies may represent p130. The nuclear accumulation of p130 may be a novel mechanism regulating nuclear β-catenin function during TMCH.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10

Similar content being viewed by others

Abbreviations

APC:

adenomatous polyposis coli

FITC:

fluorescein isothiocyanate

PAGE:

polyacrylamide gel electrophoresis

TBS:

tris buffered saline

TMCH:

transmissible murine colonic hyperplasia

References

  • Baeg GH, Matumine A, Kuroda T, Bhattacharjee RN, Miyashiro L, Yoyoshima K and Akiyama T . (1995). EMBO J., 14, 5618–5624.

  • Barthold SW . (1979). Cancer Res., 39, 24–29.

  • Barthold SW, Coleman GL, Jacoby RO, Livstone EM and Jonas AM . (1978). Vet Pathol., 15, 223–236.

  • Barthold SW and Jonas AM . (1977). Cancer Res., 37, 4352–4360.

  • Barthold SW, Osbaldiston GW and Jonas AM . (1977). Lab. Animal Sci., 27, 938–945.

  • Behrens J, Jerchow BA, Wurtele M, Grimm J, Asbrand C, Wirtz R, Kuhl M, Wedlich D and Birchmeier W . (1998). Science, 280, 596–599.

  • Brocardo M, Nathke IS and Henderson BR . (2005). EMBO Rep., 6, 184–190.

  • Browne SJ, MacFarlane M, Cohen GM and Paraskeva C . (1998). Cell Death Differ., 5, 206–213.

  • Deka J, Herter P, Sprenger-Haussels M, Koosch S, Franz D, Muller KM, Kuhnen C, Hoffmann I and Muller O . (1999). Oncogene, 18, 5654–5661.

  • Efstathiou JA, Noda M, Rowan A, Dixon C, Chinery R, Jawhari A, Hattori T, Wright NA, Bodmer WF and Pignatelli M . (1998). Proc. Natl. Acad. Sci. USA, 95, 3122–3127.

  • Eleftheriou A, Yoshida M and Henderson BR . (2001). J. Biol. Chem., 276, 25883–25888.

  • Fagotto F, Gluck U and Gumbiner BM . (1998). Curr. Biol., 8, 181–190.

  • Fearon ER and Vogelstein B . (1990). Cell, 61, 759–767.

  • Galea M, Eleftheriou A and Henderson BR . (2001). J. Biol. Chem., 276, 45833–45839.

  • Hart MJ, de los Santos R, Albert IN, Rubinfeld B and Polakis P . (1998). Curr. Biol., 8, 8:573–8:581.

  • Henderson BR . (2000). Nat. Cell Biol., 2, 653-660.

  • Horii A, Nakatsuru S, Ichii S, Nagase H and Nakamura Y . (1993). Hum. Mol. Genet., 2, 283–287.

  • Iwamoto M, Ahnen DJ, Franklin WA and Maltzman TH . (2000). Carcinogenesis, 21, 1935-1940.

  • McCartney BM and Peifer M . (2000). Nat. Cell Biol., 2, 58-60.

  • Miyashiro I, Senda T, Matsumine A, Baeg GH, Kuroda T, Shimano T, Miura S, Noda T, Kobayashi S, Monden M, Toyoshima K and Akiyama T . (1995). Oncogene, 11, 89-96.

  • Morin PJ, Vogelstein B and Kinzler KW . (1996). Proc. Natl. Acad. Sci. USA, 93, 7950–7954.

  • Munemitsu S, Alber T, Souza B, Rubinfeld B and Polakis P . (1995). Proc. Natl. Acad. Sci. USA, 92, 3046–3050.

  • Munemitsu S, Souza B, Muller O, Albert I, Rubinfeld B and Polakis P . (1994). Cancer Res., 54, 3676–3681.

  • Nakamura T, Hamada F, Ishidate T, Anai K, Kawahara K, Toyoshima K and Akiyama T . (1998). Genes Cells, 3, 395–402.

  • Nathke IS, Adam CL, Polakis P, Sellin JH and Nelson WJ . (1996). J. Cell Biol., 134, 165–179.

  • Neufeld KL and White RL . (1997). Proc. Natl. Acad. Sci. USA, 94, 3034–3039.

  • Neufeld KL, Nix DA, Bogerd H, Kang Y, Beckerle MC, Cullen BR and White RL . (2000a). Proc. Natl. Acad. Sci. USA, 97, 12085–12090.

  • Neufeld KL, Zhang F, Cullen BR and White RL . (2000b). EMBO Rep., 1, 519–523.

  • Newman JV, Kosaka T, Sheppard BJ, Fox JG and Schauer DB . (2001). J. Infect. Dis., 184, 227-230.

  • Oshima M, Sugiyama H, Kitagawa K and Taketo M . (1993). Cancer Res., 53, 5589–5591.

  • Reinacher-Schick A and Gumbiner BM . (2001). J. Cell. Biol., 152, 491-502.

  • Rosin-Arbesfeld R, Townsley F and Bienz M . (2000). Nature, 406, 1009-1012.

  • Roura S, Martinez D, Piedra J, Miravet S, Garcia de Herreros A and Dunach M . (2003). Biochem. Biophys. Res. Commun., 309, 830–835.

  • Rubinfeld B, Albert I, Porifi E, Fiol C, Munemitsu S and Polakis P . (1996). Science, 272, 1023–1026.

  • Rubinfeld B, Souza B, Alber I, Muller O, Chamberlain SH, Masiarz FR, Munemitsu S and Polakis P . (1993). Science, 262, 1731–1734.

  • Rubinfeld B, Souza B, Albert I, Munemitsu S and Polakis P . (1995). J. Biol. Chem., 270, 5549–5555.

  • Sansom OJ, Reed KR, Hayes AJ, Ireland H, Brinkmann H, Newton IP, Batlle E, Simon-Assmann P, Clevers H, Nathke IS, Clarke AR and Winton DJ . (2004). Genes Dev., 18, 1385–1390.

  • Santoro IM and Groden J . (1997). Cancer Res., 57, 488–494.

  • Sellin JH, Umar S, Xiao J and Morris AP . (2001). Cancer Res., 61, 2899–2906.

  • Smits R, Kielman MF, Breukel C, Zurcher C, Neufeld K, Jagmohan-Changur S, Hofland N, van Dijk J, White R, Edelmann W, Kucherlapati R, Khan PM and Fodde R . (1999). Genes Dev., 13, 1309-1321.

  • Spink EK, Fridman SG and Weis WI . (2001). EMBO J., 20, 6203–6212.

  • Su LK, Vogelstein B and Kinzler KW . (1993a). Science, 262, 1734–1737.

  • Thliveris A, Samowitz W, Matsunami N, Groden J and White R . (1994). Cancer Res., 54, 2991–2995.

  • Umar S, Morris AP, Kourouma F and Sellin JH . (2003). Cell Prolif., 36, 361–375.

  • Umar S, Scott J, Sellin JH, Dubinsky WP and Morris AP . (2000a). Am. J. Physiol., 278, G753–G764.

  • Umar S, Sellin JH and Morris AP . (2000b). Am. J. Physiol., 278, G765–G774.

  • Umar S, Sellin JH and Morris AP . (2000c). Am. J. Physiol., 279, G223–G237.

  • Vogelstein B, Fearon ER, Hamilton SR, Kern SE, Preisinger AC, Leppert M, Nakamura Y, White R, Smits AM and Bos JL . (1988). N. Engl. J. Med., 319, 25–532.

  • Vogelstein B and Kinzler KW . (1994). Cold Spring Harbor Symp. Quant. Biol., 59, 517–521.

  • Webb SJ, Nicholson D, Bubb VJ and Wyllie AH . (1999). FASEB J., 13, 339-346.

  • White RL . (1997). Pathol. Biol., 45, 240-244.

  • Wiechens N and Fagotto F . (2001). Curr. Biol., 11, 18–27.

  • Willert K and Nusse R . (1998). Curr. Opin. Genet. Dev., 8, 95–102.

  • Wong MH, Hermiston ML, Syder AJ and Gordon JI . (1996). Proc. Natl. Acad. Sci. USA, 93, 9588-9593.

  • Zhang F, White RL and Neufeld KL . (2000). Proc. Natl. Acad. Sci. USA, 97, 12577–12582.

  • Zhang F, White RL and Neufeld KL . (2001). Mol. Cell. Biol., 21, 8143–8156.

Download references

Acknowledgements

The authors wish to thank Dr. Andrew P Morris and Dr Leoncio Vergara for their assistance with confocal microscopy and for helpful discussion. We also thank Mr Famourou Kourouma for technical support. This work was supported by grants CA 099121 of the National Cancer Institute to SU, and Cancer Research Foundation of America to JHS, and by funds from Gastrointenstinal Research Interdisciplinary Program (GRIP), UTMB, Galveston, TX.

Author information

Authors and Affiliations

  1. Department of Internal Medicine, Division of Gastroenterology and Hepatology, University of Texas Medical Branch, 301 University Blvd, 1108 Strand, Galveston, 77555-0632, TX, USA

    Shahid Umar, Yu Wang & Joseph H Sellin

Authors
  1. Shahid Umar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joseph H Sellin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shahid Umar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Umar, S., Wang, Y. & Sellin, J. Epithelial proliferation induces novel changes in APC expression. Oncogene 24, 6709–6718 (2005). https://doi.org/10.1038/sj.onc.1208820

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1208820

Keywords

This article is cited by

Search

Advanced search

Quick links