Cyclooxygenase-2 expression in human colon cancer cells increases metastatic potential.
Journal: 1997/May - Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
PUBMED: 9096394
Abstract:
Recent epidemiologic studies have shown a 40-50% reduction in mortality from colorectal cancer in individuals who take nonsteroidal antiinflammatory drugs on a regular basis compared with those not taking these agents. One property shared by all of these drugs is their ability to inhibit cyclooxygenase (COX), a key enzyme in the conversion of arachidonic acid to prostaglandins. Two isoforms of COX have been characterized, COX-1 and COX-2. COX-2 is expressed at high levels in intestinal tumors in humans and rodents. Human colon cancer cells (Caco-2) were permanently transfected with a COX-2 expression vector or the identical vector lacking the COX-2 insert. The Caco-2 cells, which constitutively expressed COX-2, acquired increased invasiveness compared with the parental Caco-2 cells or the vector transfected control cells. Biochemical changes associated with this phenotypic change included activation of metalloproteinase-2 and increased RNA levels for the membrane-type metalloproteinase. Increased invasiveness and prostaglandin production were reversed by treatment with sulindac sulfide, a known COX inhibitor. These studies demonstrate that constitutive expression of COX-2 can lead to phenotypic changes that alter the metastatic potential of colorectal cancer cells.
Relations:
Content
Citations
(267)
References
(35)
Clinical trials
(2)
Diseases
(1)
Conditions
(2)
Drugs
(1)
Chemicals
(6)
Organisms
(3)
Processes
(1)
Anatomy
(3)
Affiliates
(1)
Similar articles
Articles by the same authors
Discussion board
Proc Natl Acad Sci U S A 94(7): 3336-3340

Cyclooxygenase-2 expression in human colon cancer cells increases metastatic potential

Departments of Medicine and Cell Biology, Molecular Toxicology Center, Vanderbilt University Medical Center and the Veterans Affairs Medical Center, Nashville, TN 37232; and First Department of Medicine, Osaka University School of Medicine, Osaka, Japan
To whom reprint requests should be addressed at: Department of Medicine/GI; MCN C-2104, Vanderbilt University Medical Center, Nashville, TN 37232-2279. e-mail: ude.tlibrednav.xavrtc@nrsiobud.
Philip Needleman, Monsanto Company, St. Louis, MO
Received 1996 Sep 18; Accepted 1997 Jan 24.

Abstract

Recent epidemiologic studies have shown a 40–50% reduction in mortality from colorectal cancer in individuals who take nonsteroidal antiinflammatory drugs on a regular basis compared with those not taking these agents. One property shared by all of these drugs is their ability to inhibit cyclooxygenase (COX), a key enzyme in the conversion of arachidonic acid to prostaglandins. Two isoforms of COX have been characterized, COX-1 and COX-2. COX-2 is expressed at high levels in intestinal tumors in humans and rodents. Human colon cancer cells (Caco-2) were permanently transfected with a COX-2 expression vector or the identical vector lacking the COX-2 insert. The Caco-2 cells, which constitutively expressed COX-2, acquired increased invasiveness compared with the parental Caco-2 cells or the vector transfected control cells. Biochemical changes associated with this phenotypic change included activation of metalloproteinase-2 and increased RNA levels for the membrane-type metalloproteinase. Increased invasiveness and prostaglandin production were reversed by treatment with sulindac sulfide, a known COX inhibitor. These studies demonstrate that constitutive expression of COX-2 can lead to phenotypic changes that alter the metastatic potential of colorectal cancer cells.

Keywords: invasion, metalloproteinase/prostaglandins/sulindac sulfide
Abstract

Colorectal cancer is the second leading cause of death from cancer in the United States. Even though this disease is curable in early stages, frequently the tumor becomes metastatic by the time an individual presents to their physician with symptoms and thus, the mortality is very high. Therefore, increasing efforts are being focused on developing more effective screening and prevention measures for colorectal cancer. Several studies have reported a 40–50% decrease in mortality from colorectal cancer in persons who are continuous users of aspirin and other nonsteroidal antiinflammatory drugs (NSAIDs) (16), suggesting that these drugs may provide a chemoprotective effect. Other studies have shown that the NSAID sulindac is effective in causing regression of adenoma size and number in patients with familial adenomatous polyposis (710). Most NSAIDs currently in use inhibit both cyclooxygenase (COX)-1 and COX-2 at their recommended dosages (1114). Two COX isoforms have been characterized, COX-1 and COX-2. COX-1 is expressed in normal intestine, but its levels do not change in intestinal tumors; however, COX-2 is undetectable in normal intestine (15) and its levels are elevated in up to 85% of colorectal adenocarcinomas (1618). We have reported that COX-2 mRNA and protein levels are elevated in colonic tumors that develop in rodents following carcinogen treatment (19) and in intestinal adenomas taken from Min mice (20). To examine the role of COX-2 in intestinal epithelial tumorigenesis we have shown that constitutive COX-2 expression in nontransformed rat intestinal epithelial cells leads to inhibition of programmed cell death (21).

The high mortality associated with colorectal cancer is related to its ability to spread beyond the large intestine and invade distant sites. Thus, tumor cell invasion is an extremely important factor for formation of solid tumors and necessary for their spread to distant organs. Proteolysis of basement membrane is one of the most important steps for invasion. Basement membrane consists of laminin, fibronectin, type IV collagen and proteoglycan. Among these basement membrane components, proteolysis of type IV collagen seems most important for invasion to occur because the network fibers of basement membrane are composed of type IV collagen.

The aim of the present study was to investigate the effect of constitutive COX-2 expression in human colon cancer cells on the invasive potential of these cells. Caco-2 cells normally express barely detectable levels of the COX-2 protein, even following growth stimulation. We found that the Caco-2 cells, programmed to constitutively expressed COX-2, acquired increased invasiveness compared with the parental Caco-2 cells or the vector transfected control cells. Biochemical changes associated with this phenotypic change included activation of membrane metalloproteinase-2 (MMP-2) and increased RNA levels for the membrane-type metalloproteinase-1 (MT-MMP-1). The increased invasiveness and prostaglandin production was reversed by treatment with sulindac sulfide, a known COX inhibitor. Our results indicate a direct link between COX-2 and activation of matrix degrading metalloproteinase enzymes.

Acknowledgments

We would like to thank Drs. H. Sato and M. Seiki for generously providing us with the MT-MMP-1 cDNA. We acknowledge support from the A. B. Hancock, Jr., Memorial Laboratory, the Lucille P. Markey Charitable Trust, the U.S. Public Health Service (Grants DK 47297 to R.N.D. and NIHES 00267 to R.N.D.), and a Veterans Affairs Merit grant (to R.N.D.). R.N.D. is a recipient of a Veterans Affairs Research Associate career development award, Boehringer Ingelheim New Investigator Award, and is an American Gastroenterological Association Gastroenterology Young Investigator Award recipient.

Acknowledgments

ABBREVIATIONS

COXcyclooxygenase
NSAIDnonsteroidal antiinflammatory drug
MMP-2metalloproteinase-2
MT MMPmembrane-type metalloproteinase
PGE2prostaglandin E2
TIMPtissue inhibitors of metalloproteinases
ABBREVIATIONS

References

  • 1. Thun M J, Namboodiri M M, Heath C W J. N Engl J Med. 1991;325:1593–1596.[PubMed]
  • 2. Thun M J, Namboodiri M M, Calle E E, Flanders W D, Heath C W J. Cancer Res. 1993;53:1322–1327.[PubMed]
  • 3. Marnett L J. Cancer Res. 1992;52:5575–5589.[PubMed]
  • 4. Marnett L J. Prev Med. 1995;24:103–106.[PubMed]
  • 5. Giovannucci E, Rimm E B, Stampfer M J, Colditz G A, Ascherio A, Willett W C. Ann Intern Med. 1994;121:241–246.[PubMed]
  • 6. Giovannucci E, Egan K M, Hunter D J, Stampfer M J, Colditz G A, Willett W C, Speizer F E. N Eng J Med. 1995;333:609–614.[PubMed]
  • 7. Waddell W R, Loughry R W. J Surg Oncol. 1983;24:83–87.[PubMed]
  • 8. Waddell W R, Gasner G F, Cerise E J, Loughry R W. Am J Surg. 1989;157:175–178.[PubMed]
  • 9. Giardiello F M, Hamilton S R, Krush A J, Piantadosi S, Hylind L M, Celano P, Booker S V, Robinson C R, Offerhaus G J. N Engl J Med. 1993;328:1313–1316.[PubMed]
  • 10. Giardiello F M, Offerhaus G J A, DuBois R N. Eur J Cancer. 1995;31A:1071–1076.[PubMed]
  • 11. Meade E A, Smith W L, DeWitt D L. J Biol Chem. 1993;268:6610–6614.[PubMed]
  • 12. Laneuville O, Breuer D K, Dewitt D L, Hla T, Funk C D, Smith W L. J Pharmacol Exp Ther. 1994;271:927–934.[PubMed]
  • 13. O’Neill G P, Mancini J A, Kargman S, Yergey J, Kwan M Y, Falgueyret J P, Abramovitz M, Kennedy B P, Ouellet M, Cromlish W, Culp S, Evans J F, Ford-Hutchinson A W, Vickers P J. Mol Pharmacol. 1994;45:245–254.[PubMed]
  • 14. Gierse J K, Hauser S D, Creely D P, Koboldt C, Rangwala S H, Isakson P C, Seibert K. Biochem J. 1995;305:479–484.
  • 15. Kargman S, Charleson S, Cartwright M, Frank J, Mancini J, Evans J, O’Neill G. Gastroenterology. 1996;111:445–454.[PubMed]
  • 16. Eberhart C E, Coffey R J, Radhika A, Giardiello F M, Ferrenbach S, DuBois R N. Gastroenterology. 1994;107:1183–1188.[PubMed]
  • 17. Kargman S, O’Neill G, Vickers P, Evans J, Mancini J, Jothy S. Cancer Res. 1995;55:2556–2559.[PubMed]
  • 18. Sano H, Kawahito Y, Wilder R L, Hashiramoto A, Mukai S, Asai K, Kimura S, Kato H, Kondo M, Hla T. Cancer Res. 1995;55:3785–3789.[PubMed]
  • 19. DuBois R N, Radhika A, Reddy B S, Entingh A J. Gastroenterology. 1996;110:1259–1262.[PubMed]
  • 20. Williams C W, Luongo C, Radhika A, Zhang T, Lamps L W, Nanney L B, Beauchamp R D, DuBois R N. Gastroenterology. 1996;111:1134–1140.[PubMed]
  • 21. Tsujii M, DuBois R N. Cell. 1995;83:493–501.[PubMed]
  • 22. DuBois R N, Tsujii M, Bishop P, Awad J A, Makita K, Lanahan A. Am J Physiol. 1994;266:G822–G827.[PubMed]
  • 23. DuBois R N, Shao J, Sheng H, Tsujii M, Beauchamp R D. Cancer Res. 1996;56:733–737.[PubMed]
  • 24. Heussen C, Dowdle E B. Anal Biochem. 1980;102:196–202.[PubMed]
  • 25. Alexander C M, Werb Z. J Cell Biol. 1992;118:727–739.
  • 26. Behrendtsen O, Alexander C M, Werb Z. Development (Cambridge, UK) 1992;114:447–456.[PubMed]
  • 27. Chirgwin J M, Przybyla A E, MacDonald R J, Rutter W J. Biochemistry. 1979;18:5294–5299.[PubMed]
  • 28. Albini A, Iwamoto Y, Kleinman H K, Martin G R, Aaronson S A, Kozlowski J M, McEwan R N. Cancer Res. 1987;47:3239–3245.[PubMed]
  • 29. Melchiori A, Albini A, Ray J M, Stetler-Stevenson W G. Cancer Res. 1992;52:2353–2356.[PubMed]
  • 30. Kobayashi H, Ohi H, Sugimura M, Shinohara H, Fujii T, Terao T. Cancer Res. 1992;52:3610–3614.[PubMed]
  • 31. Cao J, Sato H, Takino T, Seiki M. J Biol Chem. 1995;270:801–805.[PubMed]
  • 32. Strongin A Y, Collier I, Bannikov G, Marmer B L, Grant G A, Goldberg G I. J Biol Chem. 1995;270:5331–5338.[PubMed]
  • 33. Denhardt D T, Feng B, Edwards D R, Cocuzzi E T, Malyankar U M. Pharmacol Ther. 1993;59:329–341.[PubMed]
  • 34. Nomura H, Sato H, Seiki M, Mai M, Okada Y. Cancer Res. 1995;55:3263–3266.[PubMed]
  • 35. Takino T, Sato H, Shinagawa A, Seiki M. J Biol Chem. 1995;270:23013–23020.[PubMed]
Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.