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 Article
  • Published:

Activation of NF-κB is required for mediating proliferative and antiapoptotic effects of progastrin on proximal colonic crypts of mice, in vivo

Oncogene volume 27, pages 5599–5611 (2008)Cite this article

Abstract

Mice overexpressing progastrin (PG) in intestinal mucosa (fatty acid-binding protein (Fabp)-PG mice) are at an increased risk of proximal colon carcinogenesis in response to azoxymethane. Here, we report a significant increase in the length of proximal colonic crypts in Fabp-PG mice, associated with potent antiapoptotic effects of PG, which likely contributed to the previously reported increase in colon carcinogenesis in Fabp-PG mice. Phosphorylation of kinase of IκBα (IKKα/β), inhibitor of κB (IκB)α and p65NF-κB was significantly elevated in proximal colonic crypts of Fabp-PG versus wild-type mice, which was associated with degradation of IκBα and nuclear translocation/activation of p65. Surprisingly, distal colonic crypt cells were not as responsive to elevated levels of PG in Fabp-PG mice. Annexin II, recently described as a high-affinity receptor for PG, strongly co-localized with PG intracellularly and on basolateral membranes of proximal crypt cells, providing evidence that annexin-II binds PG in situ in colonic crypt cells. Proliferative and antiapoptotic effects of PG on proximal crypts of Fabp-PG mice were attenuated to wild-type levels, on treatment with NEMO peptide (an inhibitor of nuclear factor-κB (NF-κB) activation), demonstrating for the first time a critical role of NF-κB in mediating hyperproliferative affects of PG on colonic crypts of Fabp-PG mice, in vivo. Thus, downregulation of NF-κB may significantly reduce the increased risk of colon carcinogenesis in response to PG.

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

Similar content being viewed by others

Abbreviations

ANX-II:

annexin-II

IκB:

inhibitor of κB

IKK:

IκB kinase

NF-κB:

nuclear factor-κB

PG:

progastrin

PI:

proliferation index

WT:

wild-type littermates

Tg:

transgenic

WB:

western blot

References

  • Ahmed S, Murphy RF, Lovas S . (2005). Importance of N- and C-terminal regions of gastrin-Gly for preferential binding to high and low affinity gastrin-Gly receptors. Peptides 26: 1207–1212.

    Article  CAS  Google Scholar 

  • Aly A, Shulkes A, Baldwin GS . (2001). Short term infusion of glycine-extended gastrin(17) stimulates both proliferation and formation of aberrant crypt foci in rat colonic mucosa. Int J Cancer 94: 307–313.

    Article  CAS  Google Scholar 

  • Baldwin GS, Hollande F, Yang Z, Karelina Y, Paterson A, Strang R et al. (2001). Biologically active recombinant human progastrin(6-80) contains a tightly bound calcium ion. J Biol Chem 276: 7791–7796.

    Article  CAS  Google Scholar 

  • Chicone L, Narayan S, Townsend Jr CM, Singh P . (1989). The presence of a 33–40 KDa gastrin binding protein on human and mouse colon cancer. Biochem Biophys Res Commun 164: 512–519.

    Article  CAS  Google Scholar 

  • Chung CY, Erickson HP . (1994). Cell surface annexin II is a high affinity receptor for the alternatively spliced segment of tenascin-C. J Cell Biol 126: 539–548.

    Article  CAS  Google Scholar 

  • Cobb S, Wood T, Ceci J, Varro A, Velasco M, Singh P . (2004). Intestinal expression of prograstin significantly increases colon carcinogenesis in transgenic mice in response to AOM. Cancer 100: 1311–1323.

    Article  CAS  Google Scholar 

  • Dasgupta S, Jana M, Zhou Y, Fung YK, Ghosh S, Pahan K . (2004). Antineuroinflammatory effect of NF-kappaB essential modifier-binding domain peptides in the adoptive transfer model of experimental allergic encephalomyelitis. J Immunol 173: 1344–1354.

    Article  CAS  Google Scholar 

  • Dockray GJ, Varro A, Dimaline R . (1996). Gastric endocrine cells: gene expression, processing, and targeting of active products. Physiol Rev 76: 767–798.

    Article  CAS  Google Scholar 

  • Ferrand A, Bertrand C, Portolan G, Cui G, Carlson J, Pradayrol L . (2005). Signaling pathways associated with colonic mucosa hyperproliferation in mice overexpressing gastrin precursors. Cancer Res 65: 2770–2777.

    Article  CAS  Google Scholar 

  • Fric P, Sovova V, Sloncova E, Lojda Z, Jirasek A, Cermak J . (2000). Different expression of some molecular markers in sporadic cancer of the left and right colon. Eur J Cancer Prev 9: 265–268.

    Article  CAS  Google Scholar 

  • Glebov OK, Rodriguez LM, Nakahara K, Jenkins J, Cliatt J, Humbyrd CJ . (2003). Distinguishing right from left colon by the pattern of gene expression. Cancer Epidemiol Biomarkers Prev 12: 755–762.

    CAS  PubMed  Google Scholar 

  • Hajjar KA, Jacovina AT, Chacko J . (1994). An endothelial cell receptor for plasminogen/tissue plasminogen activator. I. Identity with annexin II. J Biol chem 269: 21191–21197.

    CAS  PubMed  Google Scholar 

  • Hayden MS, Ghosh S . (2004). Signaling to NF-kappa B. Genes Dev 18: 2195–2224.

    Article  CAS  Google Scholar 

  • Hong MY, Chapkin RS, Wild CP, Morris JS, Wang N, Carroll RJ et al. (1999). Relationship between DNA adduct levels, repair enzyme, and apoptosis as a function of DNA methylation by azoxymethane. Cell Growth Differ 10: 749–758.

    CAS  PubMed  Google Scholar 

  • Iacopetta B . (2002). Are there two sides to colorectal cancer? Int J Cancer 101: 403–408.

    Article  CAS  Google Scholar 

  • Johnson LR, McCormack SA . (1994). Physiology of the Gastrointestinal Tract, 3rd edn. Raven Press: New York.

    Google Scholar 

  • Lifshitz S, Lamprecht SA, Benharroch D, Prinsloo I, Polak-Charcon S, Schwartz B . (2001). Apoptosis (programmed cell death) in colonic cells: from normal to transformed stage. Cancer Lett 163: 229–238.

    Article  CAS  Google Scholar 

  • Liu LU, Holt PR, Krivosheyev V, Moss SF . (1999). Human right and left colon differ in epithelial cell apoptosis and in expression of Bak, a pro-apoptotic Bcl-2 homologue. Gut 45: 45–50.

    Article  CAS  Google Scholar 

  • Marshman E, Ottewell PD, Potten CS, Watson AJ . (2001). Caspase activation during spontaneous and radiation-induced apoptosis in the murine intestine. J Pathol 195: 285–292.

    Article  CAS  Google Scholar 

  • May MJ, D'Acquisto F, Madge LA, Glöckner J, Pober JS, Ghosh S . (2000). Selective inhibition of NF-kappaB activation by a peptide that blocks the interaction of NEMO with the IkappaB kinase complex. Science 289: 1550–1554.

    Article  CAS  Google Scholar 

  • Mills SJ, Mathers JC, Chapman PD, Burn J, Gunn A . (2001). Colonic crypt cell proliferation state assessed by whole crypt microdissection in sporadic neoplasia and familial adenomatous polyposis. Gut 48: 41–46.

    Article  CAS  Google Scholar 

  • Müerköster S, Isberner A, Arlt A, Witt M, Reimann B, Blaszczuk E et al. (2005). Gastrin suppresses growth of CCK2 receptor expressing colon cancer cells by inducing apoptosis in vitro and in vivo. Gastroenterology 129: 952–968.

    Article  Google Scholar 

  • Ortiz-Zapater E, Peiró S, Roda O, Corominas JM, Aguilar S, Ampurdanés C et al. (2007). Tissue plasminogen activator induces pancreatic cancer cell proliferation by a non-catalytic mechanism that requires extracellular signal-regulated kinase 1/2 activation through epidermal growth factor receptor and annexin A2. Am J Pathol 170: 1573–1584.

    Article  CAS  Google Scholar 

  • Ottewell PD, Varro A, Dockray GJ, Kirton CM, Watson AJ, Wang TC et al. (2005). COOH-terminal 26-amino acid residues of progastrin are sufficient for stimulation of mitosis in murine colonic epithelium in vivo. Am J Phsyiol Gastrointest Liver Physiol 288: G541–G549.

    Article  CAS  Google Scholar 

  • Ottewell PD, Watson AJ, Wang TC, Varro A, Dockray GJ, Pritchard DM . (2003). Progastrin stimulates murine colonic epithelial mitosis after DNA damage. Gastroenterology 124: 1348–1357.

    Article  CAS  Google Scholar 

  • Rengifo-Cam W, Singh P . (2004). Role of progastrins and gastrins and their receptors in GI and pancreatic cancers: targets for treatment. Curr Pharm Des 10: 2345–2358.

    Article  CAS  Google Scholar 

  • Rengifo-Cam W, Umar S, Sarkar S, Singh P . (2007). Anti-apoptotic effects of progastrin on pancreatic cancer cells are mediated by sustained activation of nuclear factor κB. Cancer Res 67: 7266–7274.

    Article  CAS  Google Scholar 

  • Sarkar S, Umar S, Singh G, Singh P . (2007). Progastrin (PG) peptide co-localizes with annexin II (ANX-II), in situ, in human colon cancer cells (HCT-116), expressing autocrine PG, and in intestinal epithelial cells (IEC-18) in response to PG stimulation. Gastroenterology 132: A540–A541.

    Google Scholar 

  • Sato M, Ahnen DJ . (1992). Regional variability of colonocyte growth and differentiation in the rat. Anat Rec 233: 409–414.

    Article  CAS  Google Scholar 

  • Seva C, Dickinson CJ, Yamada T . (1994). Growth-promoting effects of glycine-extended progastrin. Science 265: 410–412.

    Article  CAS  Google Scholar 

  • Singh P . (2007). Role of annexin-II in GI cancers: interaction with gastrins/progastrins. Cancer Lett 252: 19–35.

    Article  CAS  Google Scholar 

  • Singh P, Lu X, Cobb S, Miller BT, Tarasova N, Varro A et al. (2003). Progastrin1-80 stimulates growth of intestinal epithelial cells in vitro via high-affinity binding sites. Am J Physiol Gastrointest Liver Physiol 284: G328–G339.

    Article  CAS  Google Scholar 

  • Singh P, Velasco M, Given R, Varro A, Wang TC . (2000A). Progastrin expression predisposes mice to colon carcinomas and adenomas in response to a chemical carcinogen. Gastroenterology 119: 162–171.

    Article  CAS  Google Scholar 

  • Singh P, Velasco M, Given R, Wargovich M, Varro A, Wang TC . (2000B). Mice overexpressing progastrin are predisposed for developing aberrant colonic crypt foci in response to AOM. Am J Physiol Gastrointest Liver Physiol 278: G390–G399.

    Article  CAS  Google Scholar 

  • Singh P, Wu H, Clark C, Owlia A . (2007). Annexin II binds progastrin and gastrin-like peptides, and mediates growth factor effects of autocrine and exogenous gastrins on colon cancer and intestinal epithelial cells. Oncogene 26: 425–440.

    Article  CAS  Google Scholar 

  • Song DH, Rana B, Wolfe JR, Crimmins G, Choi C, Albanese C . (2003). Gastrin-induced gastric adenocarcinoma growth is mediated through cyclin D1. Am J Physiol Gastrointest Liver Physiol 285: G217–G222.

    Article  CAS  Google Scholar 

  • Umar S, Morris AP, Kourouma F, Sellin JH . (2003). Dietary pectin and calcium inhibit colonic proliferation in vivo by differing mechanisms. Cell Prolif 36: 361–375.

    Article  CAS  Google Scholar 

  • Umar S, Sellin JH, Morris AP . (2000). Increased nuclear translocation of catalytically active PKC-zeta during mouse colonocyte hyperproliferation. Am J Physiol Gastrointest Liver Physiol 279: G223–G237.

    Article  CAS  Google Scholar 

  • Umar S, Wang Y, Morris AP, Sellin JH . (2007). Dual alterations in casein kinase I-epsilon and GSK-3beta modulate beta-catenin stability in hyperproliferating colonic epithelia. Am J Physiol Gastrointest Liver Physiol 292: G599–G5607.

    Article  CAS  Google Scholar 

  • Wang TC, Koh TJ, Varro A, Cahill RJ, Dangler CA, Fox JG et al. (1996). Processing and proliferative effects of human progastrin in transgenic mice. J Clin Invest 98: 1918–1929.

    Article  CAS  Google Scholar 

  • Wang Y, Xiang GS, Kourouma F, Umar S . (2006). Citrobacter rodentium-induced NF-kappa B activation in hyperproliferating colonic epithelia: role of p65 (Ser(536)) phosphorylation. Br J Pharmacol 148: 814–824.

    Article  CAS  Google Scholar 

  • Watanabe T, Kobunai T, Toda E, Yamamoto Y, Kanazawa T, Kazama Y . (2006). Distal colorectal cancers with microsatellite instability (MSI) display distinct gene expression profiles that are different from proximal MSI cancers. Cancer Res 66: 9804–9808.

    Article  CAS  Google Scholar 

  • Wu H, Owlia A, Singh P . (2003). Precursor peptide progastrin(1-80) reduces apoptosis of intestinal epithelial cells and upregulates cytochrome c oxidase Vb levels and synthesis of ATP. Am J Physiol Gastrointest Liver Physiol 285: G1097–G1110.

    Article  CAS  Google Scholar 

  • Zizzo MG, Mule F, Serio R . (2006). Inhibitory responses to exogenous adenosine in murine proximal and distal colon. Br J Pharmacol 148: 956–963.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grant nos. CA97959 and CA114264 to P Singh, and Grant no. CA099121 to S Umar from the NIH.

Author information

Author notes

  1. S Umar and S Sarkar: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX, USA

    S Umar

  2. Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX, USA

    S Sarkar, S Cowey & P Singh

Authors
  1. S Umar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S Sarkar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Cowey
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P Singh.

Additional information

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Umar, S., Sarkar, S., Cowey, S. et al. Activation of NF-κB is required for mediating proliferative and antiapoptotic effects of progastrin on proximal colonic crypts of mice, in vivo. Oncogene 27, 5599–5611 (2008). https://doi.org/10.1038/onc.2008.169

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2008.169

Keywords

This article is cited by

Search

Advanced search

Quick links