Skip to main content
SpringerLink
Log in

Glycine-Extended Gastrin Exerts Growth-Promoting Effects on Human Colon Cancer Cells

  • Original Articles
  • Published:
Molecular Medicine Aims and scope Submit manuscript

Abstract

Background

Since human colon cancers often contain significant quantities of progastrin-processing intermediates, we sought to explore the possibility that the biosynthetic precursor of fully processed amidated gastrin, glycine-extended gastrin, may exert trophic effects on human colonic cancer cells.

Materials and Methods

Binding of radiolabeled glycine-extended and amidated gastrins was assessed on five human cancer cell lines: LoVo, HT 29, HCT 116, Colo 320DM, and T 84. Trophic actions of the peptides were assessed by increases in [3H]thymidine incorporation and cell number. Gastrin expression was determined by northern blot and radioimmunoassay.

Results

Amidated gastrin did not bind to or stimulate the growth of any of the five cell lines. In contrast, saturable binding of radiolabeled glycine-extended gastrin was seen on LoVo and HT 29 cells that was not inhibited by amidated gastrin (10−6 M) nor by a gastrin/CCKB receptor antagonist (PD 134308). Glycine-extended gastrin induced a dose-dependent increase in [3H]thymidine uptake in LoVo (143 ± 8% versus control at 10−10 M) and HT 29 (151 ± 11% versus control at 10−10 M) cells that was not inhibited by PD 134308 or by a mitogen-activated protein (MAP) or ERK kinase (MEK) inhibitor (PD 98509). Glycine-extended gastrin did stimulate jun-kinase activity in LoVo and HT 29 cells. The two cell lines expressed the gastrin gene at low levels and secreted small amounts of amidated gastrin and glycine-extended gastrin into the media.

Conclusions

Glycine-extended gastrin receptors are present on human colon cancer cells that mediate glycine-extended gastrin’s trophic effects via a MEK-independent mechanism. This suggests that glycine-extended gastrin and its novel receptors may play a role in colon cancer cell growth.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  1. Johnson LR. (1977) New aspects of the trophic action of gastrointestinal hormones. Gastroenterology 72: 788–792.

    PubMed  CAS  Google Scholar 

  2. Rehfeld JF. (1995) Gastrin and colorectal cancer: A never-ending dispute? Gastroenterology 108: 1307–1309.

    Article  CAS  PubMed  Google Scholar 

  3. Watson SA, Durrant LG, Crosbie JD, Morris DL. (1989) The in vitro growth response of primary human colorectal and gastric cancer cells to gastrin. Int. J. Cancer 43: 692–696.

    Article  CAS  PubMed  Google Scholar 

  4. McGregor DB, Jones RD, Karlin DA, Romsdahl MM. (1989) Trophic effects of gastrin on colorectal neoplasms in the rat. Ann. Surg. 195: 219–223.

    Article  Google Scholar 

  5. Winsett OE, Townsend CM, Glass EJ, Thompson JC. (1986) Gastrin stimulates growth of colon cancer. Surgery 99: 302–307.

    PubMed  CAS  Google Scholar 

  6. Tatsuta M, Yamamura H, Iishi H, Noguchi S, Ichii M, and Tangiuchi H. (1985) Gastrin has no promoting effect on chemically induced colonic tumors in Wistar rats. Eur. J. Cancer Clin. Oncol 21: 741–744.

    Article  CAS  PubMed  Google Scholar 

  7. Oscarson JEA, Veen HF, Ross JS, Malt RA. (1982) Dimethylhydrazine induced colonic neoplasia: dissociation from endogenous gastrin levels. Surgery 91: 525–530.

    PubMed  CAS  Google Scholar 

  8. Parsonnet J, Kim P, Yang S, Orentreich JH, Friedmann GD. (1996) Gastrin and gastric adenocarcinoma: a prospective evaluation. Gastroenterology 110: A574.

    Google Scholar 

  9. Thorburn CM Freidman GD, Dickinson CJ, Vogelman JH, Orentreich N, Parsonnet J. (1998) Gastrin and colorectal cancer: a prospective study. Gastroenterology 115: 275–280.

    Article  CAS  PubMed  Google Scholar 

  10. Kochman ML, Del Valle J, Dickinson CJ, Boland CR. (1992) Post-translational processing of gastrin in neoplastic human colonic tissue. Biochem. Biophys. Res. Commun. 189: 1165–1169.

    Article  CAS  PubMed  Google Scholar 

  11. Nemeth J, Taylor B, Pauwels S, Varro A, Dockray GJ. (1993) Identification of progastrin derived peptides in colorectal carcinoma extracts. Gut 34: 90–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Ciccotosto GD, McLeish A, Hardy KJ, Shulkes A. (1995) Expression, processing and secretion of gastrin in patients with colorectal carcinoma. Gastroenterology 109: 1142–1153.

    Article  CAS  PubMed  Google Scholar 

  13. van Solinge WW, Neilsen FC, Friis-Hansen L, Falkmer U, Rehfeld JF. (1993) Expression but incomplete processing of progastrin in colorectal carcinomas. Gastroenterology 104: 1099–1107.

    Article  PubMed  Google Scholar 

  14. Merchant JL, Dickinson CJ, Yamada T. (1994) Molecular biology of the gut: model of gastrointestinal hormones. In: Johnson LR (ed). Physiology of the Gastrointestinal Tract, 3rd ed. Raven Press, New York, pp. 295–350.

    Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Rosenquist GL, Walsh JH. (1979) Radioimmunoassay of gastrin. In: Jerzy Glass GB (ed). Gastrointestinal Hormones. Raven Press, New York, pp 769–795.

    Google Scholar 

  18. Todisco A, Takeuchi Y, Seva C, Dickinson CJ, Yamada T. (1995) Gastrin and glycine-extended processing intermediates induce different programs of early gene activation. J. Biol. Chem. 270: 28337–28341.

    Article  CAS  PubMed  Google Scholar 

  19. Hibi M, Lin A, Smeal T, Minden A, Karin M. (1993) Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev. 7: 2135–2148.

    Article  CAS  PubMed  Google Scholar 

  20. Guan KL, Dixon JE. (1991) Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione 5-transferase. Anal. Biochem. 192: 262–267.

    Article  CAS  PubMed  Google Scholar 

  21. Boel E, Vuust J, Norris F, Wind A, Rehfeld JF, Marcker KA. (1983) Molecular cloning of human gastrin cDNA: evidence for evolution of gastrin by gene duplication. Proc. Natl. Acad. Sci. U.S.A. 80: 2866–2869.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Wilborg O, Berglund L, Boel E, et al. (1984) Structure of a human gastrin gene. Proc. Natl. Acad. Sci. U.S.A. 81: 1067–1069.

    Article  Google Scholar 

  23. Marino LR, Muglia BH, Dickinson CJ. (1994) Glycine-extended post-translational processing intermediates of gastrin and cholecystokinin in the gut. Regul. Pept. 50: 73–85.

    Article  CAS  PubMed  Google Scholar 

  24. Todisco A, Yamada J, Uromov A, Yamada T. (1996) Molecular mechanisms for the growth factor action of gastrin. J. Invest. Med. 44: 318A.

    Google Scholar 

  25. Johnson LR. (1977) Gastrointestinal hormones and their functions. Annu. Rev. Physiol. 39: 135–158.

    Article  CAS  PubMed  Google Scholar 

  26. Kusyk CJ, McNiel NO, Johnson LR. (1986) Stimulation of growth of a colon cancer cell line by gastrin. Am. J. Physiol. 251: G597–G601.

    Article  CAS  PubMed  Google Scholar 

  27. Smith JP, Stock EA, Wotring MG, McLaughlin PJ, Zagon IS. (1996) Characterization of the CCK-B/gastrin-like receptor in human colon cancer. Am. J. Physiol. 271: R797–R805.

    PubMed  CAS  Google Scholar 

  28. Winsett OE, Townsend CM Jr, Glass EJ, Thompson CJ. (1986) Gastrin stimulates growth of colon cancer. Surgery 99: 302–307.

    PubMed  CAS  Google Scholar 

  29. Chu M, Nielsen FC, Franzen L, Rehfeld JF, Holst JJ, Borch K. (1995) Effect of endogenous hypergastrinemia on gastrin receptor expressing human colon carcinoma transplanted to athymic rats. Gastroenterology 109: 1415–1420.

    Article  CAS  PubMed  Google Scholar 

  30. Dickinson CJ. (1995) Relationship of gastrin processing to colon cancer. Gastroenterology 109: 1384–1388.

    Article  CAS  PubMed  Google Scholar 

  31. Singh P, Xu Z, Dai B, Rajaraman S, Rubin N, Dhruva B. (1994) Incomplete processing of progastrin expressed by human colon cancer cells: role of noncarboxyamidated gastrins. Am. J. Physiol. 266: G459–G468.

    PubMed  CAS  Google Scholar 

  32. Rehfeld JF, Hansen HF. (1986) Characterization of preprocholecystokinin products in the porcine cerebral cortex: evidence of different processing pathways. J. Biol. Chem. 261: 5832–5840.

    PubMed  CAS  Google Scholar 

  33. Sugano K, Aponte GW, Yamada T. (1985) Identification and characterization of glycine-extended post-translational processing intermediates of progastrin in porcine stomach. J. Biol. Chem. 260: 11724–11729.

    PubMed  CAS  Google Scholar 

  34. DelValle J, Sugano K, Yamada T. (1987) Progastrin and its glycine-extended posttranslational processing intermediates in human gastrointestinal tissues. Gastroenterology 92: 1908–1912.

    Article  CAS  Google Scholar 

  35. Sugano K, Park J, Dobbins WO, Yamada T. (1987) Glycine-extended progastrin processing intermediates: Accumulation and cosecretion with gastrin. Am. J. Physiol. 253: G502–G507.

    PubMed  CAS  Google Scholar 

  36. DelValle J, Sugano K, Yamada T. (1989) Glycine-extended processing intermediates of gastrin and cholecystokinin in human plasma. Gastroenterology 97: 1159–1163.

    Article  CAS  PubMed  Google Scholar 

  37. Jensen S, Borch K, Hilsted L, Rehfeld JF. (1989) Progastrin processing during antral G-cell hypersecretion in humans. Gastroenterology 96: 1063–1070.

    Article  CAS  PubMed  Google Scholar 

  38. Hilsted L. (1991) Glycine-extended gastrin precursors. Regul. Pept. 36: 323–343.

    Article  CAS  PubMed  Google Scholar 

  39. Singh P, Owlia A, Espenijo R, Dai B. (1995) Novel gastrin receptors mediate mitogenic effects of gastrin and processing intermediates of gastrin on Swiss 3T3 fibroblasts. J. Biol. Chem. 270: 8429–8438.

    Article  CAS  PubMed  Google Scholar 

  40. Bold RJ, Ishizuka J, Townsend CMJ, Thompson JC. (1994) Gastrin stimulates growth of human colon cancer cells via a receptor other than CCK-A or CCK-B. Biochem. Biophys. Res. Commun. 202: 1222–1226.

    Article  CAS  PubMed  Google Scholar 

  41. Koh TJ, Dockray GJ, Varro A, Cahill RJ, Fox JG, Wang TC. (1997) Overexpression of glycine-extended progastrin results in gastric ulceration and goblet cell hyperplasia. Gastroenterology 112: A1163.

    Article  Google Scholar 

  42. Hilsted L, Hint K, Christiansen J, Rehfeld JF. (1988) Neither glycine-extended, nor the 1–13 fragment of gastrin 17 influences gastric acid secretion in humans. Gastroenterology 94: 96–110.

    Article  CAS  PubMed  Google Scholar 

  43. Matsumoto M, Park J, Sugano K, Yamada T. (1987) Biological activity of progastrin posttranslational processing intermediates. Am. J. Physiol. 87: G315–G319.

    Google Scholar 

  44. Singh P, Owlia A, Varro A, Dai BS, Rajaraman S, Wood T. (1996) Gastrin gene expression is required for the proliferation and tumorigenicity of human colon cancer cells. Cancer Res. 56: 4111–4115.

    PubMed  CAS  Google Scholar 

  45. Miyake A. (1995) A truncated isoform of human CCK-B/gastrin receptor generated by alternative usage of a novel exon. Biochem. Biophys. Res. Commun. 208: 230–237.

    Article  CAS  PubMed  Google Scholar 

  46. Hoosein NM, Kiener PA, Curry RC, Rovati LC, McGilbra DK, Brattain MG. (1988) Antiproliferative effects of gastrin receptor antagonists and antibodies to gastrin on human colon carcinoma cell lines. Cancer Res. 48: 7179–7183.

    PubMed  CAS  Google Scholar 

  47. Blackmore M, Hirst BH. (1992) Autocrine stimulation of growth of AR 42J rat pancreatic tumor cells by gastrin. Br. J. Cancer 66: 32–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Scemama JL, Fourmy D, Zahidi A, Pradayrol L, Susini C, Ribet A. (1987) Characterization of gastrin receptors on a rat pancreatic acinar cell line (AR42J). Gut 28 (Suppl): 233–236.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Davis JR. (1993) The mitogen-activated protein kinase signal transduction pathway. J. Biol. Chem. 268: 14553–14556.

    PubMed  CAS  Google Scholar 

  50. Marais R, Wynne J, Treisman R. (1993) The SRF accessory protein Elk-1 contains a growth factor-regulated transcriptional activation domain. Cell 73: 381–393.

    Article  CAS  PubMed  Google Scholar 

  51. Treisman R. (1995) Journey to the surface of the cell: Fos regulation and the SRE. EMBO J. 14: 4905–4913.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Alessi DR, Cuenda A, Cohen P, Dudley DT, Saltiel A. (1995) PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J. Biol. Chem. 270: 27489–27494.

    Article  CAS  PubMed  Google Scholar 

  53. Cobb MH, Goldsmith EJ. (1995) How MAP kinases are regulated. J. Biol. Chem. 270: 14843–14846.

    Article  CAS  PubMed  Google Scholar 

  54. Clark GJ, Westwick JK, Der CJ. (1997) p120 GAP modulates Ras activation of jun kinases and transformation. J. Biol. Chem. 272: 1677–1681.

    Article  CAS  PubMed  Google Scholar 

  55. Minden A, Lin A, Smeal T, Derijard B, Cobb M, Davis R, Karin M. (1994) c-Jun N-terminal phosphorylation correlates with activation of the JNK subgroup of mitogen-activated protein kinases. Mol. Cell Biol. 14: 6683–6688.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Whitmarsh AJ, Shore P, Sharrocks AD, Davis RJ. (1995) Integration of MAP kinase signal transduction pathways at the serum response element. Science 269: 403–407.

    Article  CAS  PubMed  Google Scholar 

  57. Cavigelli M, Dolfi F, Claret FX, Karin M. (1995) Induction of c-fos expression through JNK-mediated TCF/Elk-1 phosphorylation. EMBO J. 14: 5957–5964.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Westwick JK, Weitzel C, Minden A, Karin M, Brenner DA. (1994) Tumor necrosis factor a stimulates AP-1 activity through prolonged activation of the c-jun kinase. J. Biol. Chem. 269: 26396–26401.

    PubMed  CAS  Google Scholar 

  59. Westwick JK, Bielawska AE, Dbaibo G, Hannun YA, Brenner DA. (1995) Ceramide activates the stress-activated protein kinases. J. Biol. Chem. 270: 22689–22692.

    Article  CAS  PubMed  Google Scholar 

  60. Licato LL, Keku TO, Sandler RS, Brenner DA. (1996) JNK and ERK kinases are activated in DMH-induced colonic tumors. Gastroenterology 110: A551.

    Google Scholar 

  61. Higashide S, Gomez G, Greeley GH Jr, Townsend CM Jr, Thompson JC. (1996) Glycine-extended gastrin potentiates gastrin-stimulated acid secretion in rats. Am. J. Physiol. 270: G220–G224.

    PubMed  CAS  Google Scholar 

  62. Kaise M, Muraoka A, Seva C, Takeda H, Dickinson CJ, Yamada T. (1995) Glycine-extended progastrin intermediates induce H+,K+-ATPase a-sub-unit gene expression through a novel receptor. J. Biol. Chem. 270: 1155–11160.

    Google Scholar 

  63. Negre F, Fagot-Revurat P, Bouisson M, Rehfeld JF, Vaysse N. (1995) Autocrine stimulation of AR4-2J rat pancreatic tumor cell growth by glycine-extended gastrin. Int. J. Cancer 66: 653–658.

    Article  Google Scholar 

  64. Varro A, Voronina S, Dockray GJ. (1995) Pathways of processing of the gastrin precursor in rat antral mucosa. J. Clin. Invest. 95: 1642–1649.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported in part by funds from the National Institutes of Health grants RO1DK34306, RO1DK47398, and KO8DK02336 (A. T.) and by funds from the University of Michigan Gastrointestinal Peptide Research Center (National Institutes of Health grant P30DK34933). A. T. is the recipient of an American Gastroenterological Association Industry Research Scholar Award. V. S. has received funding from a Max Kade Postdoctoral Research Grant (Max Kade Foundation, New York, NY).

Author information

Authors and Affiliations

  1. Department of Pediatric Gastroenterology, University of Michigan, Medical Center, 1150 West Medical Center Drive, A520C MSRB I, Ann Arbor, Michigan, 48109-0658, USA

    Vinzenz M. Stepan, Mitsutaka Sawada &  Chris J. Dickinson

  2. Department of Internal Medicine, University of Michigan, Medical Center, Ann Arbor, Michigan, USA

    Vinzenz M. Stepan, Mitsutaka Sawada & Andrea Todisco

Authors
  1. Vinzenz M. Stepan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mitsutaka Sawada
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrea Todisco
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chris J. Dickinson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chris J. Dickinson.

Additional information

Communicated by T. Yamada.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stepan, V.M., Sawada, M., Todisco, A. et al. Glycine-Extended Gastrin Exerts Growth-Promoting Effects on Human Colon Cancer Cells. Mol Med 5, 147–159 (1999). https://doi.org/10.1007/BF03402058

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03402058

Keywords

Search

Navigation