Pharmaceutical Research

, 25:2151 | Cite as

Chemoprevention of Colon Carcinogenesis by Oleanolic Acid and Its Analog in Male F344 Rats and Modulation of COX-2 and Apoptosis in Human Colon HT-29 Cancer Cells

  • Naveena B. Janakiram
  • Cooma Indranie
  • Swamy V. Malisetty
  • Patlolla Jagan
  • Vernon E. Steele
  • Chinthalapally V. Rao
Research Paper



To assess the chemopreventive effect of oleanolic acid (ONA) and its synthetic analog 18α-olean-12-ene-3β-23,28-triol (OT) on azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) in F344 rats and understand anti-inflammatory properties and apoptosis effects in HT29 colon cancer cells and Raw 264.7 macrophage cell lines.


Five week-old male F344 rats were fed a control diet or experimental diets containing two doses of ONA (750 and 1,500 ppm) and OT (250 and 500 ppm). After 1 week, all animals were s.c. injected with AOM (15 mg/kg body weight, once weekly for 2 weeks). At 14 weeks of age, all rats were killed and colons were evaluated for ACF. Cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) expressions and apoptosis were assessed in cell lines exposed to OT using western blots and 4’,6-diamidino-2-phenylindole staining.


Administration of ONA and OT inhibited mean colonic ACF and multi-crypt AC/foci in a dose dependent manner (p < 0.001–0.0001). OT blocked the COX-2 expression induced by phorbol 12-myristate 13-acetate in a dose-dependent manner and induced apoptosis in HT-29 cancer cells, and suppressed iNOS activation in RAW264.7 macrophages.


ONA and OT possess chemopreventive activity against colon carcinogenesis in rat and OT inhibits the COX-2 and iNOS and induces apoptosis in cell lines.

Key words

chemoprevention colon cancer COX-2 iNOS triterpenoids 



We gratefully acknowledge NCI chemoprevention drug development repository for providing oleanolic acid analog OT. We also extend our thanks to Ms. Alyson Atchison for editing this article.


  1. 1.
    S. B. Mahato, and S. Sen. Advances in triterpenoid research 1990–1994. Phytochemistry. 44:1185–1236 (1997).PubMedCrossRefGoogle Scholar
  2. 2.
    N. P. Sahu, S. B. Mahato, S. K. Sarkar, and G. Poddar. Triterpenoid saponins from Gymnema sylvestre. Phytochemistry. 41:1181–1185 (1996).PubMedCrossRefGoogle Scholar
  3. 3.
    J. Liu. Pharmacology of oleanolic acid and ursolic acid. J. Ethnopharmacol. 49:57–68 (1995).PubMedCrossRefGoogle Scholar
  4. 4.
    N. Suh, Y. Wang, T. Honda, G. W. Gribble, E. Dmitrovsky, W. F. Hickey, R. A. Maue, A. E. Place, D. M. Porter, M. J. Spinella, C. R. Williams, G. Wu, A. J. Dannenberg, K. C. Flanders, J. J. Letterio, D. J. Mangelsdorf, C. F. Nathan, L. Nguyen, W. W. Porter, R. F. Ren, N. S. Roche, K. Subbaramaiah, and M. B. Sporn. A novel synthetic oleanane triterpenoid, 2-cyno-3,12-dioxoolean-1,9-dien-28-oic acid, with potent differentiating antiproliferative and anti-inflammatory activity. Cancer Res. 59:336–341 (1999).PubMedGoogle Scholar
  5. 5.
    B. J. Naveena, and C. V. Rao. Molecular markers and targets for colorectal cancer prevention. Acta Pharmacologica Sinica. 29:1–20 (2008).Google Scholar
  6. 6.
    C. V. Rao. Nitric oxide signaling in colon cancer chemoprevention. Mutat. Res. 555:107–119 (2004).PubMedGoogle Scholar
  7. 7.
    C. V. Rao, C. Indranie, B. Simi, P. T. Manning, J. R. Connor, and B. S. Reddy. Chemopreventive properties of a selective inducible nitric oxide synthase inhibitor in colon carcinogenesis, administered alone or in combination with celecoxib, a selective cyclooxygenase-2 inhibitor. Cancer Res. 62(1):165–70 (2002).PubMedGoogle Scholar
  8. 8.
    M. Takahashi, K. Fukuda, T. Ohata, T. Sugimura, and K. Wakabayashi. Increased expression of inducible and endothelial constitutive nitric oxide synthases in rat colon tumors induced by azoxymethane. Cancer Res. 57:1233–1237 (1997).PubMedGoogle Scholar
  9. 9.
    M. Tsujii, S. Kawano, S. Tsuji, H. Sawaoka, M. Hori, and R. Dubois. Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell. 93:705–716 (1998).PubMedCrossRefGoogle Scholar
  10. 10.
    T. Hida, H. A. Yatabe, H. Muramatsu, K. Kozaki, S. Nakamura, M. Ogawa, M. Mitsudomi, S. Iwata, and S. Shibata. Increased expression of cyclooxygenase 2 occurs frequently in human lung cancers, specifically in adenocarcinomas. Cancer Res. 58:3761–3764 (1998).PubMedGoogle Scholar
  11. 11.
    R. P. Bird. Role of aberrant crypt foci in understanding the pathogenesis of colon cancer. Cancer Lett. 93:55–71 (1995).PubMedCrossRefGoogle Scholar
  12. 12.
    P. Greenwald, C. K. Clifford, and J. A. Milner. Diet and cancer prevention. Eur. J. Cancer. 37:948–965 (2001).PubMedCrossRefGoogle Scholar
  13. 13.
    T. Fung, F. B. Hu, C. Fuchs, E. Giovannucci, D. J. Hunter, M. J. Stampfer, G. A. Colditz, and W. C. Willet. Major dietary patterns and the risk of colorectal cancer in women. Arch. Intern. Med. 163:309–314 (2003).PubMedCrossRefGoogle Scholar
  14. 14.
    B. Chiu, B. Ji, Q. Dai, G. Gridley, J. K. Mclaughlin, Y. Gao, J. F. Fraumeni, and W. Chow. Dietary factors and risk of colon cancer in Shanghai, China. Cancer Epidemiol. Biomarkers Prev. 12:201–208 (2003).PubMedGoogle Scholar
  15. 15.
    H. Nishino, A. Nishino, J. Takayasu, T. Hasegawa, A. Iwashima, K. Hirabayashi, S. Iwata, and S. Shibata. Inhibition of the tumor-promoting action of 12-O-tetradecanoylphorbol-13-acetate by some oleanane-type triterpenoid compounds. Cancer Res. 48:5210–5215 (1988).PubMedGoogle Scholar
  16. 16.
    G. B. Singh, S. Singh, S. Bani, B. D. Gupta, and S. K. Banerjee. Anti-inflammatory activity of oleanolic acid in rats and mice. J. Pharm. Pharmacol. 44:456–458 (1992).PubMedGoogle Scholar
  17. 17.
    M. T. Huang, C. T. Ho, Z. Y. Wang, T. Ferraro, Y. R. Lou, K. Stauber, W. Ma, C. Georgiadis, J. D. Laskin, and A. H. Conney. Inhibition of skin tumorigenesis by rosemary and its constituents carnosol and ursolic acid. Cancer Res. 54:701–708 (1994).PubMedGoogle Scholar
  18. 18.
    C. Indranie, S. V. Malisetty, P. Jagan, L. E. S. Joel, V. E. Steele, and C. V. Rao. Chemoprevention of colon carcinogenesis by oleanolic acid and its analog in male F344rats and modulation of iNOS, COX-2 and apoptosis in human colon HT-29 cancer cells. AACR Proceeding. Abstract No. 819. 43:164–164 (2002).Google Scholar
  19. 19.
    C. V. Rao, B. Simi, and B. S. Reddy. Inhibition by dietary curcumin of azoxymethane-induced ornithine decarboxylase, tyrosine protein kinase, arachidonic acid metabolism and aberrant crypt foci formation in the rat colon. Carcinogenesis. 14(11):2219–2225 (1993).PubMedCrossRefGoogle Scholar
  20. 20.
    C. V. Rao, D. Desai, B. Simi, N. Kulkarni, S. Amin, and B. S. Reddy. Inhibitory effect of caffeic acid esters on azoxymethane-induced biochemical changes and aberrant crypt foci formation in rat colon. Cancer Res. 53:4182–4188 (1993).PubMedGoogle Scholar
  21. 21.
    B. J. Naveena, C. Indranie, M. Altaf, V. E. Steele, and C. V. Rao. b-ionone inhibits colonic aberrant crypt foci in rats, suppresses cell growth and induces retinoid X receptor a in human colon cancer cells. Mol. Cancer Ther. 7(1):181–190 (2008).CrossRefGoogle Scholar
  22. 22.
    R. Jayadev, M. R. P. Jagan, V. S. Malisetty, and C. V. Rao. Colon cancer preventive effects of Trigonella foenum graecum (fenugreek) seed and its constituent diosgenin in vivo and in vitro. Cancer Epidemiol. Biomarkers Prev. 13:1392–1398 (2004).Google Scholar
  23. 23.
    T. Kawamoori, T. Tanaka, A. Hara, J. Yamahara, and H. Mori. Modifying effects of naturally occurring products on the development of colonic aberrant crypt foci by azoxymethane in F344 rats. Cancer Res. 55:1277–1282 (1995).Google Scholar
  24. 24.
    H. Ohigashi, H. Takamura, K. Koshimizu, H. Tokuda, and Y. Ito. Search for possible antitumor promoters by inhibition of n-O-tetradecanoylphorbol-B-acetate induced Epstein–Barr virus activation: ursolic acid and oleanolic acid from an anti-inflammatory Chinese medicinal plant, Glechoma hederaceae L. Cancer Lett. JO:143–151 (1986).CrossRefGoogle Scholar
  25. 25.
    N. Yoshimi, A. Wang, Y. Morishita, T. Tanaka, S. Sugie, K. Kawai, J. Yamahara, and H. Mori. Modifying effects of fungal and herb metabolites on azoxymethane-induced intestinal carcinogenesis in rats. Jpn. J. Cancer Res. 83:1273–1278 (1992).PubMedGoogle Scholar
  26. 26.
    Z. Ovesna, A. Vachalkova, K. Horvathova, and D. Tothova. Pentacyclic triterpenic acids: New chemoprotective compounds. Neoplasma. 51:327–333 (2004).PubMedGoogle Scholar
  27. 27.
    Gu. Yongchuan, G. Wang, G. Pan, J. P. Fawcett, A. Jiye, and S. Jianguo. Transport and bioavailability studies of astragaloside IV, an active ingredient in radix astragali. Basic Clin. Pharmacol. Toxicol. 95(6):295–298 (2004).CrossRefGoogle Scholar
  28. 28.
    Y. Motoo, and N. Sawabu. Antitumor effects of saikosaponins, baicalin and baicalein on human hepatoma cell lines. Cancer Lett. 86:91–95 (1994).PubMedCrossRefGoogle Scholar
  29. 29.
    H. Yano, A. Mizoguchi, K. Fukuda, M. Haramaki, S. Ogasawara, S. Momosaki, and M. Kojiro. The herbal medicine sho-saiko-to inhibits proliferation of cancer cell lines by inducing apoptosis and arrest at the G0/G1 phase. Cancer Res. 54:448–454 (1994).PubMedGoogle Scholar
  30. 30.
    L. Qian, T. Murakami, Y. Kimura, M. Takahashi, and K. Okita. Saikosaponin A-induced cell death of a human hepatoma cell line (HuH-7): the significance of the ‘sub-G1 peak’ in a DNA histogram. Pathol. Int. 45:207–214 (1995).PubMedCrossRefGoogle Scholar
  31. 31.
    M. B. Sporn, and A. B. Roberts. Peptide growth factors and inflammation, tissue repair, and cancer. J. Clin. Investig. 78:329–332 (1986).PubMedCrossRefGoogle Scholar
  32. 32.
    H. Ohshima, and H. Bartsch. Chronic infections and inflammatory processes as cancer risk factors: possible role of nitric oxide in carcinogenesis. Mutat. Res. 305:253–264 (1994).PubMedGoogle Scholar
  33. 33.
    G. Steinbach, P. M. Lynch, R. K. Phillips, M. H. Wallace, E. Hawk, G. B. Gordon, N. Wakabayashi, B. Saunders, Y. Shen, T. Fujimura, L. K. Su, and B. Levin. The effect of celecoxib, a cyclooxygenase-2 inhibitor, in familial adenomatous polyposis. N. Engl. J. Med. 342:1946–1952 (2000).PubMedCrossRefGoogle Scholar
  34. 34.
    K. Muller-Decker, G. Neufang, I. Berger, M. Neumann, F. Marks, and G. Furstenberger. Transgenic cyclooxygenase-2 overexpression sensitizes mouse skin for carcinogenesis. Proc. Natl. Acad. Sci. 99:12483–12488 (2002).PubMedCrossRefGoogle Scholar
  35. 35.
    M. E. Alvarez, A. E. Rotelli, L. E. Pelzer, J. R. Saad, and O. Giordano. Phytochemical study and anti-inflammatory properties of Lampaya hieronymi Schum. ex Moldenke II Farmaco. 55(6–7):502–505 (2000).CrossRefGoogle Scholar
  36. 36.
    H. Xiangjiu, and R. H. Liu. Triterpenoids isolated from apple peels have potent antiproliferative activity and may be partially responsible for apple’s anticancer activity. J. Agric. Food Chem. 55(11):4366–4370 (2007).CrossRefGoogle Scholar
  37. 37.
    T. L. Karen, M. Y. Mark, and M. B. Sporn. Triterpenoids and rexinoids as multifunctional agents for the prevention and treatment of cancer. Nat. Rev. Cancer. 7:357–369 (2007).CrossRefGoogle Scholar
  38. 38.
    S. D. Solomon, J. J. V. McMurray, M. A. Pfeffer, J. Wittes, R. Fowler, P. Finn, W. F. Anderson, A. Zauber, E. Hawk, and M. Bertagnolli. Cardiovascular risk associated with celecoxib in a clinical trial for colorectal adenoma prevention. N. Engl. J. Med. 352:1071–1080 (2005).PubMedCrossRefGoogle Scholar
  39. 39.
    A. Rostom, C. Dubé, G. Lewin, A. Tsertsvadze, N. Barrowman, and C. Code. Nonsteroidal anti-inflammatory drugs and cyclooxygenase-2 inhibitors for primary prevention of colorectal cancer: a systematic review prepared for the U.S. preventive services task force. Ann. Intern. Medicine. 146:376–89 (2007).Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Naveena B. Janakiram
    • 1
  • Cooma Indranie
    • 1
  • Swamy V. Malisetty
    • 1
  • Patlolla Jagan
    • 1
  • Vernon E. Steele
    • 2
  • Chinthalapally V. Rao
    • 1
    • 3
  1. 1.Department of Medicine, Hem–Onc Section, OU Cancer InstituteUniversity of Oklahoma Health Sciences CenterOklahoma CityUSA
  2. 2.Chemopreventive Agent Development Research Group, Division of Cancer PreventionNCI, NIHBethesdaUSA
  3. 3.OU Cancer Institute, OUHSCOklahoma CityUSA

Personalised recommendations