Inhibition of Azoxymethane Initiated Colon Tumor and Aberrant Crypt Foci Development by Bovine Lactoferrin Administration in F344 Rats

  • Hiroyuki Tsuda
  • Kazunori Sekine
  • Joe Nakamura
  • Yoshihiko Ushida
  • Tetsuya Kuhara
  • Nobuo Takasuka
  • Dae Joong Kim
  • Makoto Asamoto
  • Hiroyasu Baba-Toriyama
  • Malcolm A. Moore
  • Hoyoku Nishino
  • Tadao Kakizoe
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 443)


The influence of bovine lactoferrin (bLf) on colon carcinogenesis was investigated in male F344 rats treated with azoxymethane (AOM). In experiment I, 2% and 0.2% bLf, and Bifidobacterium longum (B. longum) as a positive control at 3% were given in the diet for 4 weeks, along with two s.c. 15 mg/kg injections of AOM on days 1 and 8. The numbers of aberrant crypt foci (ACF) were decreased by both treatments. Similar results were obtained in experiment II of 13 weeks duration. In experiment III, animals were given three weekly injections of AOM and then received 2 or 0.2% bLf, 2% bLf-hydrolysate, or 0.1% bovine lactoferricin (bLfcin) for 36 weeks. No effects indicative of toxicity were noted, but significant reduction in both the incidence and number of adenocarcinomas of the large intestine was observed with almost all the treatments. Thus, the incidences of colon adenocarcinomas in the groups receiving 2 or 0.2% bLf, 2% bLf-hydrolysate, or 0.1% bLfcin were 15%, 25%, 26.3% and only 10%, respectively, in contrast to the 57.5% control value (p < 0.01). ACF values also exhibited reduced development. Investigation of beta-glucuronidase revealed decrease in the cecal contents of animals receiving bLf. In addition, demonstration of enhancement of NK activity by bLf indicated that its inhibitory effects could have been related to elevated immune cytotoxicity.


Whey Protein Basal Diet Aberrant Crypt Focus Colon Carcinogenesis Cecal Content 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Giovannucci, E. and Willett, W. C. Dietary factors and risk of colon cancer. Ann. Med., 26: 443–452, 1994.PubMedCrossRefGoogle Scholar
  2. 2.
    Wynder, E. L., Fujita, Y., Harris, R. E., Hirayama, T. and Hiyama, T. Comparative epidemiology of cancer between the United States and Japan, a second look. Cancer, 67: 746–63, 1991.PubMedCrossRefGoogle Scholar
  3. 3.
    Wattenberg, L. W. Inhibition of carcinogenesis by minor dietary constituents. Cancer Res. (suppl.), 52: 2085s - 2091s, 1992.Google Scholar
  4. 4.
    McIntosh, G. H., Regester, G. O., Lc, L. R., Royle, P. J. and Smithers, G. W. Dairy proteins protect against dimethylhydrazinc-induced intestinal cancers in rats. J. Nutr., 125: 809–816, 1995.PubMedGoogle Scholar
  5. 5.
    Lonncrdal, B. and lyer. S. Lactoferrin: molecular structure and biological function review. Ann. Rev. Nutr., 15: 93–110, 1995.CrossRefGoogle Scholar
  6. 6.
    Nishiya, K. and Horwitz, D. A. Contrasting effects of lactoferrin on human lymphocyte and monocyte natural killer activity and antibody-dependent cell-mediated cytotoxicity. J. Immunol., 129: 2519–2523, 1982.PubMedGoogle Scholar
  7. 7.
    Shan, H., Kim. A. and Golub, S. H. Modulation of natural killer and lymphokine-activated killer cell cytotoxicity by lactoferrin. J. Leukocyte Biol., 51: 343–349, 1992.Google Scholar
  8. 8.
    Gahr, M., Speer, C. P., Damerau, B. and Sawatzki, G. Influence of lactoferrin on the function of human polymorphonuclear leukocytes and monocytes. J. Leukocyte Biol., 49: 427–433. 1991.PubMedGoogle Scholar
  9. 9.
    Sawatzki_ G. and Rich, I. N. Lactoferrin stimulates colony stimulating factor production in vitro and in vivo. Blood Cells, 15: 371–385, 1989.PubMedGoogle Scholar
  10. 10.
    McCormick, J. A., Markey, G. M. and Morris, T. C. Lactoferrin-inducible monocyte cytotoxicity for K562 cells and decay of natural killer lymphocyte cytotoxicity. Clin. Exp. Immunol., 83: 154–156, 1991.CrossRefGoogle Scholar
  11. 11.
    Levay, P. F. and Viljoen, M. Lactoferrin: a general review. Haematologica, 80: 252–267, 1995.PubMedGoogle Scholar
  12. 12.
    Bezault, J., l3himani. R., Wiprovnick, J. and Furmanski, P. Human lactoferrin inhibits growth of solid tu-mors and development of experimental metastases in mice. Cancer Res., 54: 231–2312, 1994.Google Scholar
  13. 13.
    Reddy, B. S., Simi, B., Patel. N., Aliaga, C. and Rao, C. V. Effect of amount and types of dietary fat on intestinal bacterial 7 alpha-dehydroxylase and phosphatidylinositol-specific phospholipase C and colonic mucosal diacylglycerol kinase and PKC activities during stages of colon tumor promotion. Cancer Res., 56: 2314–2320, 1996.Google Scholar
  14. 14.
    Reddy, B. S. and Rivenson, A. Inhibitory effect of Bi/ìdohacteriurn longuet on colon, mammary, and liver carcinogenesis induced by 2-amino-3-methylimidazo[4,5-fjquinoline, a food mutagen. Cancer Res., 53: 3914–3918, 1993.PubMedGoogle Scholar
  15. 15.
    Abdelali, F1., Cassand, P., Soussotte, V., Daubeze, M., Bouley, C. and Narbonne, J. F. Effect of dairy products on initiation of precursor lesions of colon cancer in rats. Nutr. Cancer, 24: 121–132, 1995.Google Scholar
  16. 16.
    Kulkarni, N. and Reddy, B. S. Inhibitory effect of Bifidobacterium longum cultures on the azoxymethaneinduced aberrant crypt foci formation and fecal bacterial beta-glucuronidase. Proc. Soc. Exp. Biot. Med., 207: 278–283, 1994.CrossRefGoogle Scholar
  17. 17.
    Tornita, M., Bellamy, W., Takasc, M., Yamauchi, K., Wakabayashi, H. and Kawase, K. Potent antibacterial peptides generated by pepsin digestion of bovine lactoferrin. J. Dairy Sci., 74: 4137–4142, 1991.CrossRefGoogle Scholar
  18. 18.
    Bellamy, W.. Takase, M., Wakabayashi, FI., Kawase. K. and Tornita, M. Antibacterial spectrum of lactoferricin B, a potent bactericidal peptide derived from the N-terminal region of bovine lactoferrin. J. Applied Bacteriol., 73: 472 479. 1992.Google Scholar
  19. 19.
    Shimamura. S.., Abe. F., Ishibashi, N., Miyakawa, H., Yaeshima, T., Araya, T. and Tornita, M. Relationship between oxygen sensitivity and oxygen metabolism of Bifidobacterium species. J. Dairy Sei., 75: 3296 3306, 1992.Google Scholar
  20. 20.
    Winter, 13. K., Wu, S., Nelson, A. C. and Pollack. S. B. Renal cell carcinoma and natural killer cells: studies in a novel rat model in vitro and in vivo. Cancer Res., 52: 6279–6286, 1992.Google Scholar
  21. 21.
    Sekine, K., Ohta, J., Onishi, M., Tatsuki, T., Shimokawa, Y., Toida, T., Kawashima, T. and Hashimoto, Y. Analysis of antitumor properties of effector cells stimulated with a cell wall preparation (WPG) of Bifido-bacterium inl4ntis. Biol. Pharm. Bull., 18: 148–153, 1995.CrossRefGoogle Scholar
  22. 22.
    Rowland, I. R., Mallett, A. K. and Wise, A. A comparison of the activity of five microbial enzymes in cecal content fì-om rats, mice, and hamsters, and response to dietary pectin. Toxicol. Appt. Pharm., 69: 143–148, 1983.CrossRefGoogle Scholar
  23. 23.
    Tudek, 13., Bird, R. P. and Bruce, W. R. Foci of aberrant crypts in the colons of mice and rats exposed to carcinogens associated with foods. Cancer Res., 49: 1236–1240, 1989.Google Scholar
  24. 24.
    Bird. R. P., Yao, K., Lasko, C. M. and Good. C. K. Inability of low-or high-fat diet to modulate late stages of colon carcinogenesis in Sprague-Dawley rats. Cancer Res., 56: 2896–2899, 1996.Google Scholar
  25. 25.
    Kampman, E.. Goldbohm, R. A., Van den Brandt, P. A. and Van’t Veer, P. Fermented dairy products, calcium, and colorectal cancer in the Netherlands cohort study. Cancer Rcs., 54: 3186–3190, 1994.Google Scholar
  26. 26.
    Sorenson, A. W., Slattery, M. L. and Ford, M. H. Calcium and colon cancer: a review. Nutr. Cancer, 2: 135–145, 1988.CrossRefGoogle Scholar
  27. 27.
    Tsuru, S., Shinomiya, N., Taniguchi, M., Shimazaki, H., Tanigawa, K. and Nomoto, K. Inhibition of tumor growth by dairy products. J. Clin. Lab. Immunol., 25: 177–183, 1988.Google Scholar
  28. 28.
    Bounous, G.. Papenburg, R., Kongshavn, P. A., Gold, P. and Fleiszer, D. Dietary whey protein inhibits the development of dimethylhydrazine induced malignancy. Clin. Invest. Med., 2: 213 217, 1988.Google Scholar
  29. 29.
    Papenburg, R., Bounous, G., Fleiszer, D. and Gold, P. Dietary milk proteins inhibit the development of dimethylhydrazine-induced malignancy. Tumour Biol., 11: 129–136, 1990.PubMedCrossRefGoogle Scholar
  30. 30.
    Kennedy, R. S., Konok, G. P., Bounous, G., Baruchel, S. and Lee, T. D. The use of a whey protein concentrate in the treatment of patients with metastatic carcinoma: a phase 1–11 clinical study. Anticancer Res., 15: 2643–2649, 1995.PubMedGoogle Scholar
  31. 31.
    Bird, R. P. Role of aberrant crypt foci in understanding the pathogenesis of colon cancer. Cancer Lett., 93: 55–71, 1995.PubMedCrossRefGoogle Scholar
  32. 32.
    Brock, J. Lactoferrin: a multifunctional immunoregulatory protein? Immunol. Today, 16: 417–419, 1995.Google Scholar
  33. 33.
    Sekine, K., Kawashima, T. and Hashimoto, Y. Comparison of the TNF-a levels induced by human-derived Bifidobacterium longum and rat-derived Bifidobacterium animalis in mouse peritoneal cells. Bifidobacteria Microflora, 13: 79–89, 1994.Google Scholar
  34. 34.
    Wei, W. Z., Fulton, A., Winkelhake, J. and Heppner, G. Correlation of natural killer activity with tumorigenesis of a preneoplastic mouse mammary lesion. Cancer Res., 49: 2709–2715, 1989.PubMedGoogle Scholar
  35. 35.
    Bounous, G., Batist, G. and Gold, P. Whey protein in cancer prevention. Cancer Lett., 57: 91–94, 1991.PubMedCrossRefGoogle Scholar
  36. 36.
    Goldin, B. R. and Gorbach, S. L. The effect of milk and lactobacillus feeding on human intestinal bacterial enzyme activity. A. J. Clin. Nutr., 39: 756–761, 1984.Google Scholar

Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Hiroyuki Tsuda
    • 1
  • Kazunori Sekine
    • 1
  • Joe Nakamura
    • 1
  • Yoshihiko Ushida
    • 1
  • Tetsuya Kuhara
    • 1
  • Nobuo Takasuka
    • 1
  • Dae Joong Kim
    • 1
  • Makoto Asamoto
    • 1
  • Hiroyasu Baba-Toriyama
    • 1
  • Malcolm A. Moore
    • 1
  • Hoyoku Nishino
    • 2
  • Tadao Kakizoe
    • 3
  1. 1.Chemotherapy DivisionNational Cancer Center Research InstituteChuo-ku, Tokyo 104Japan
  2. 2.Department of BiochemistryKyoto Prefectural University of MedicineKamigyo-ku, Kyoto 602Japan
  3. 3.National Cancer Center Central HospitalChuo-ku, Tokyo 104Japan

Personalised recommendations