Molecular and Cellular Biochemistry

, Volume 231, Issue 1–2, pp 139–146 | Cite as

Differential modulation of transforming growth factor-βs and cyclooxygenases in the platelet lysates of male F344 rats by dietary lipids and piroxicam

  • Jayadev Raju
  • Ranjan P. Bird


Platelets are implicated in the pathogenesis of various chronic diseases including cancer. The main objective of the present study was to determine if dietary fish oil and piroxicam, known modulators of colon tumorigenesis, effect transforming growth factor (TGF)-βs and cyclooxygenase (COX) isozymes in the platelets of colon tumor-bearing male F344 rats. TGF-βs and COXs are important in the development of chronic illnesses including colon cancer. Animals harboring preneoplastic colonic lesions were randomly allocated to a low fat diet (5% by weight – low corn oil, LFC) and three high fat diets (23% by weight – high corn oil, HFC; high corn oil containing 150-ppm piroxicam, HFC+P; and high fish oil, HFF) for 16 weeks. TGF-β1, TGF-β2, COX-1 and COX-2 protein levels were assessed in the platelets by Western blot analysis. Active TGF-β1 (12.5 kDa) level was significantly lower in the platelets of the HFC+P group (p < 0.001), whereas precursor TGF-β1 (39 kDa) level was significantly lower in the platelets of the HFF group (p < 0.001). The anti-rabbit TGF-β2 polyclonal antibody did not detect the 13-kDa active TGF-β2 protein in the platelets. However a 29-kDa protein, potentially a precursor of TGF-β2, was detected in the platelets of all the groups and was significantly lower in the HFC+P and HFF groups than in LFC and HFC (p < 0.001). COX-1 level was significantly lower in the HFF group than the other three groups (p < 0.001). COX-2 protein was detected in the platelets of all diet groups. Piroxicam in the presence of high corn oil (HFC+P) significantly lowered the level of COX-2 (p < 0.001), without having any effect on COX-1 level. These findings conclusively show that LFC and HFC differ from HFF and HFC+P, and piroxicam differs from fish oil, in regulating the levels of TGF-βs and COX in the platelets. This supports the conjecture that the levels of bioactive constituents of the platelets are profoundly modulated by dietary lipids, which in turn could influence the pathogenesis of chronic illnesses.

dietary lipids fish oil piroxicam platelets transforming growth factor-β cyclooxygenase carcinogenesis 


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  1. 1.
    Gold LI: The role for transforming growth factor-beta (TGF-beta) in human cancer. Crit Rev Oncog 10: 303–360, 1999Google Scholar
  2. 2.
    Kelly DL, Rizzino A: Growth regulatory factors and carcinogenesis: The roles played by transforming growth factor beta, its receptors and signalling pathways. Anticancer Res 19: 4791–4807, 1999Google Scholar
  3. 3.
    Katori M, Majima M: Cycloooxygenase-2: Its rich diversity of roles and possible application of its selective inhibitors. Inflamm Res 49: 367–392, 2000Google Scholar
  4. 4.
    Prescott SM, Fitzpatrick FA: Cyclooxygenase-2 and carcinogenesis. Biochim Biophys Acta 1470: M69–M78, 2000Google Scholar
  5. 5.
    Kim YJ, Borsig L, Han HL, Varki NM, Varki A: Distinct selectin ligands on colon carcinoma mucins can mediate pathological interactions among platelets, leukocytes and endothelium. Am J Pathol 155: 461–472, 1999Google Scholar
  6. 6.
    Puolakkainen P, Twardzik D, Ranchalis J, Moroni M, Mandeli J, Paciucci PA: Increase of plasma transforming growth factor beta (TGF beta) during immunotherapy with IL-2. Cancer Invest 13: 583–589, 1995Google Scholar
  7. 7.
    Belloc C, Lu H, Soria C, Fridman R, Legrand Y, Menashi S: The effect of platelets on invasiveness and protease production of human mammary tumor cells. Int J Cancer 60: 413–417, 1995Google Scholar
  8. 8.
    Sobol AB, Watala C: The role of platelets in diabetes-related vascular complications. Diabetes Res Clin Pract 50: 1–16, 2000Google Scholar
  9. 9.
    George JN: Platelets. Lancet 355: 1531–1539, 2000Google Scholar
  10. 10.
    Adam JM, Raju J, Khalil N, Bird RP: Evidence for the involvement of dietary lipids on the modulation of transforming growth factor-β1 in the platelets of male rats. Mol Cell Biochem 211: 145–152, 2000Google Scholar
  11. 11.
    Loi C, Chardigny JM, Almanza S, Leclere L, Ginies C, Louis S: Incorporation and metabolism of dietary trans isomers of linolenic acid alter the fatty acid profile of rat tissues. J Nutr 130: 2550–2555, 2000Google Scholar
  12. 12.
    Saker KE, Eddy AL, Thatcher CD, Kalnitsky J Manipulation of dietary (n-6) and (n-3) fatty acids alters platelet function in cats. J Nutr 128(suppl): 2645S–2647S, 1998Google Scholar
  13. 13.
    Vognild E, Elvevoll EO, Brox J, Olsen RL, Barstad H, Aursand M, Osterud B: Effects of dietary marine oils and olive oil on fatty acid composition, platelet membrane fluidity, platelet responses and serum lipids in healthy humans. Lipids 33: 427–436, 1998Google Scholar
  14. 14.
    Assoian RK, Komoriya A, Meyers CA, Miller DM, Sporn MB: Transforming growth factor-b in human platelets: Identification of a major storage site, purification and characterization. J Biol Chem 258: 7155–7160, 1983Google Scholar
  15. 15.
    Massague J: The transforming growth factor-β family. Ann Rev Cell Biol 6: 597–641, 1990Google Scholar
  16. 16.
    Heldin CH, Miyazono K, ten Dijke P: TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature 390: 465–471, 1997Google Scholar
  17. 17.
    Burmester JK, Qian SW, Roberts AB, Huang A, Amatyakul-Chantler S, Suardet L, Odartchenko N, Madri JA, Sporn MB: Characterization of distinct functional domains of transforming growth factor β. Proc Natl Acad Sci USA 90: 8628–8632, 1993Google Scholar
  18. 18.
    Okada F, Yamguchi K, Ichihara A, Nakamura T: One of two subunits of masking protein in latent TGF-β is a part of pro-TGF-β. FEBS Lett 242: 240–244, 1989Google Scholar
  19. 19.
    Wakefield LM, Smith DM, Flanders KC, Sporn MB: Latent transforming growth factor-beta from human platelets. A high molecular weight complex containing precursor sequences. J Biol Chem 263: 7646–7654, 1988Google Scholar
  20. 20.
    Blobe GC, Schiemann WP, Lodish HF: Role of transforming growth factor beta in human disease. N Engl J Med 342: 1350–1358, 2000Google Scholar
  21. 21.
    Ravitz MJ, Wenner CE: Cyclin-dependent kinase regulation during G1 phase and cell cycle regulation by TGF-β. Adv Cancer Res 71: 165–207, 1997Google Scholar
  22. 22.
    Tsushima H, Kawata S, Tamura S, Ito N, Shirai Y, Kiso S, Imai Y, Shimomukai H, Nomura Y, Matsuda Y, Matsuzawa Y: High levels of transforming growth factor beta 1 in patients with colorectal cancer: Association with disease progression. Gastroenterology 110: 375–382, 1996Google Scholar
  23. 23.
    Taipale J, Saharinen J, Keski-Oja J: Extracellular matrix-associated transforming growth factor-beta: Role in cancer cell growth and invasion. Adv Cancer Res 75: 87–134, 1998Google Scholar
  24. 24.
    Fink SP, Swinler SE, Lutterbaugh JD, Massague J, Thiagalingam S, Kinzler KW, Vogelstein B, Willson JK, Markowitz S: Transforming growth factor-beta-induced growth inhibition in a Smad4 mutant colon adenoma cell line. Cancer Res 61: 256–260, 2001Google Scholar
  25. 25.
    Dubois RN, Abramson SB, Crofford L, Gupta RA, Simon LS, Van De Putte LB, Lipsky PE: Cyclooxygenase in biology and disease. FASEB J 12, 1063–1073, 1992Google Scholar
  26. 26.
    Smith WL: Prostanoid biosynthesis and mechanisms of action. Am J Physiol 263: F181–F191, 1992Google Scholar
  27. 27.
    Jun SS, Chen Z, Pace MC, Shaul PW: Estrogen upregulates cyclooxygenase-1 gene expression in ovine fetal pulmonary artery endothelium. J Clin Invest 102: 176–183, 1999Google Scholar
  28. 28.
    Harris RC, Wang JL, Cheng HF, Zhang MZ, McKanna JA: Prostaglandins in macula densa function. Kidney Int 67: S49–S52, 1998Google Scholar
  29. 29.
    McAdam BF, Catella-Lawson F, Mardini IA, Kapoor S, Lawson JA, FitzGerald GA: Systemic biosynthesis of prostacyclin by cyclooxygenase (COX)-2: The human pharmacology of a selective inhibitor of COX-2. Proc Natl Acad Sci USA 96: 272–277, 1999Google Scholar
  30. 30.
    Kulkarni SK, Jain NK, Singh A: Cyclooxygenase isoenzymes and newer therapeutic potential for selective COX-2 inhibitors. Meth Find Exp Clin Pharmacol 22: 291–298, 2000Google Scholar
  31. 31.
    Goodwin JS: Mechanism of action of nonsteroidal anti-inflammatory agents. Am J Med 77: 57–64, 1984Google Scholar
  32. 32.
    Patrignani P, Sciulli MG, Manarini S, Santini G, Cerletti C, Evangelista V: COX-2 is not involved in thromboxanes biosynthesis by activated human platelets. J Physiol Pharmacol 50: 661–667, 1999Google Scholar
  33. 33.
    Takhashi Y, Ueda N, Yoshimoto T, Yokoama C, Miyata A, Tanabe T, Fuse I, Hattori A, Shibata A: Immunoaffinity purification and cDNA cloning of human platelet prostaglandin endoperoxide synthase (cyclooxygenase). Biochem Biophys Res Commun 182: 433–438, 1992Google Scholar
  34. 34.
    Weber A-A, Zimmermann KC, Meyer-Kirchrath J, Schror K: Cyclooxygenase-2 in human platelets as a possible factor in aspirin resistance. Lancet 353: 900, 1999Google Scholar
  35. 35.
    Saeed SA, Afzal MN, Shah BH: Dual effects of nimesulide, a COX-2 inhibitor, in human platelets. Life Sci 63: 1835–1841, 1998Google Scholar
  36. 36.
    American Institute of Nutrition: Report of the ad hoc committee on standards for nutrition studies. J Nutr 110: 1726, 1977Google Scholar
  37. 37.
    Folch J, Lees M, Stanley GHS: A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226: 497–509, 1957Google Scholar
  38. 38.
    Heemskerk J, Feijge M, Kalafusz R, Hornstra G: Influence of dietary fatty acids on membrane fluidity and activation of rat platelets. Biochim Biophys Acta 1004: 252–260, 1989Google Scholar
  39. 39.
    Bradford M: A rapid and sensitive method for the determination of microgram quantities of protein utilizing the principles of dye binding. Anal Biochem 84: 639–641, 1976Google Scholar
  40. 40.
    Bird RP, Good CK: The significance of aberrant crypt foci in understanding the pathogenesis of colon cancer. Toxicol Lett 112-113: 395–402, 2000Google Scholar
  41. 41.
    Good CK: Regulation of the Cellular, Molecular and Morphological Determinants of Colonic Precancerous Stages by Dietary Lipids. Ph.D. Thesis, The University of Manitoba, Winnipeg, Canada, 1999, pp 114–131.Google Scholar
  42. 42.
    Nordoy A, Hatcher L, Goodnight S, Fitzgerald GA, Conner WE: Effects of dietary fat content, saturated fatty acids, and fish oil on eicosanoid production and hemostatic parameters in normal men. J Lab Clin Med 123: 914–920, 1994Google Scholar
  43. 43.
    Gilbert M, Achard F, Dalloz S, Maclouf J, Benistant C, Lagarde M: Opposite regulation of prostaglandin H synthase isoforms by eicosapentaenoic and docosahexaenoic acids. Lipids 34(suppl): S219, 1999Google Scholar
  44. 44.
    Achard F, Gilbert M, Benistant C, Ben-Slama S, DeWitt DL, Smith WL, Lagarde M: Eicosapentaenoic and docosahexaenoic acids reduce PGH synthase 1 expression in bovine aortic endothelial cells. Biochem Biophys Res Commun 241: 513–518, 1997Google Scholar
  45. 45.
    Good CK, Lasko CM, Adam JM, Bird RP: Diverse effect of fish oil on the growth of aberrant crypt foci and tumor multiplicity in F344 rats. Nut Cancer 31: 204–211, 1998Google Scholar
  46. 46.
    Ritland SR, Gendler SJ: Chemoprevention of intestinal adenomas in the ApcMin mouse by piroxicam: Kinetics, strain effects and resistance to chemosupression. Carcinogenesis 20: 51–59, 1999Google Scholar

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© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Jayadev Raju
  • Ranjan P. Bird

There are no affiliations available

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