Cancer Chemotherapy and Pharmacology

, Volume 58, Issue 4, pp 451–459 | Cite as

The tumor specific cytotoxicity of dihydronitidine from Toddalia asiatica Lam

  • Hironori Iwasaki
  • Hirosuke Oku
  • Ryo Takara
  • Hanako Miyahira
  • Kaoru Hanashiro
  • Yasuhiko Yoshida
  • Yasuhiro Kamada
  • Tetsuya Toyokawa
  • Kensaku Takara
  • Masashi Inafuku
Original Article
First Online:
Received:
Accepted:

Abstract

Purpose: In recent years, a number of reports have shown the anticancer activity of plant extracts and phytoalkaloid. Methods: We have evaluated the cytotoxicity profiles of 157 extracts prepared from dietary or medical plants growing in the Okinawa island, using 10 different cell lines. In vitro cytotoxicity screening indicated the presence of a highly selective cytotoxic compound in the extract of Toddalia asiatica Lam. The known alkaloid (1,3)benzodioxolo(5,6-c)phenanthridine, 12,13-dihydro-2,3-dimethoxy-12-methyl-(dihydronitidine) was identified as an active material from this plant. This alkaloid had highly specific cytotoxicity to human lung adenocarcinoma (A549) cells. Results: The results of the fluorescence activated cell sorter (FACS) analysis and the measurement of caspase-3 activity showed that dihydronitidine induced specific apoptotic cell death in A549 cells. Gene expression analysis in the apoptotic cells found that dihydronitidine variously regulated the cell cycle related genes (CDK2 and CCNE), and up-regulated the cell death related genes specifically in tumor cells. Thus dihydronitidine manifested its characteristics in the tumor selective cytotoxicity, contrasting with the case of a known anticancer agent camptothecin (CPT). Microscopic observation further revealed the specific accumulation of dihydronitidine within the cytosolic organelle, but not in the nuclei of adenocarcinoma. No accumulation was observed with CPT in all cell lines. Conclusion: The data suggested that dihydronitidine toxicity targeted a particular intracellular organelle in the tumor cells.

Keywords

Tumor Alkaloids Apoptosis Accumulation Cytotoxicity Dihydronitidine 
This is a preview of subscription content, log in to check access.

References

  1. 1.
    Wall ME (1998) Camptothecin and taxol: discovery to clinic. Med Res Rev 18:299–314CrossRefPubMedGoogle Scholar
  2. 2.
    Wink M (2003) Evolution of secondary metabolites from an ecological and molecular phylogenetic perspective. Phytochemistry 64:3–19CrossRefPubMedGoogle Scholar
  3. 3.
    Cai Y, Luo Q, Sun M, Corke H (2004) Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 74:2157–2184CrossRefPubMedGoogle Scholar
  4. 4.
    Kritz H, Sinzinger H (1997) Tea consumption, lipid metabolism, and atherosclerosis Wien Klin Wochenschr 109:944–948PubMedGoogle Scholar
  5. 5.
    Valcic S, Timmermann BN, Alberts DS, Wachter GA, Krutzsch M, Wymer J, Guillen JM (1996) Inhibitory effect of six green tea catechins and caffeine on the growth of four selected human tumor cell lines. Anticancer Drugs 7:461–468PubMedCrossRefGoogle Scholar
  6. 6.
    Savarese A, Cognetti F (2003) New drugs in the treatment of recurrent or metastatic cervical cancer. Crit Rev Oncol Hematol 48:323–327PubMedCrossRefGoogle Scholar
  7. 7.
    Pisani P, Parkin DM, Bray F, Ferlay J (1999) Estimates of the worldwide mortality from 25 cancers in 1990. Int J Cancer 83:18–29CrossRefPubMedGoogle Scholar
  8. 8.
    Cragg GM, Schepartz SA, Suffness M, Grever MR (1993) The taxol supply crisis New NCI policies for handling the large-scale production of novel natural product anticancer and anti-HIV agents. J Nat Prod 56:1657–1668CrossRefPubMedGoogle Scholar
  9. 9.
    Rothenberg ML (1997) Topoisomerase I inhibitors: review and update. Ann Oncol 8:837–855CrossRefPubMedGoogle Scholar
  10. 10.
    Lee JH, Lee JM, Kim JK, Ahn SK, Lee SJ, Kim MY, Jew SS, Park JG, Hong CI (1998) Antitumor activity of 7-[2-(N-isopropylamino)ethyl]-(20S)-camptothecin, CKD602, as a potent DNA topoisomerase I inhibitor. Arch Pharm Res 21:581–590PubMedCrossRefGoogle Scholar
  11. 11.
    Lee JH, Lee JM, Lim KH, Kim JK, Ahn SK, Bang YJ, Hong CI (2000) Preclinical and phase I clinical studies with Ckd-602, a novel camptothecin derivative. Ann N Y Acad Sci 922:324–325PubMedCrossRefGoogle Scholar
  12. 12.
    Vanhoefer U, Harstrick A, Achterrath W, Cao S, Seeber S, Rustum YM (2001) Irinotecan in the treatment of colorectal cancer: clinical overview. J Clin Oncol 19:1501–1518PubMedGoogle Scholar
  13. 13.
    Gore M, ten BokkelHuininkW, Carmichael J, Gordon A, Davidson N, Coleman R, Spaczynski M, Heron JF, Bolis G, Malmstrom H, Malfetano J, Scarabelli C, Vennin P, Ross G, Fields SZ (2001) Clinical evidence for topotecan-paclitaxel non–cross-resistance in ovarian cancer. J Clin Oncol 19:1893–1900PubMedGoogle Scholar
  14. 14.
    Oketch-Rabah HA, Mwangi JW, Lisgarten J, Mberu EK (2000) A new antiplasmodial coumarin from Toddalia asiatica roots. Fitoterapia 71:636–640CrossRefPubMedGoogle Scholar
  15. 15.
    Kshirsagar RD, Singh NP (2001) Some less known ethnomedicinal uses from Mysore and Coorg districts, Karnataka state, India. J Ethnopharmacol 75:231–238CrossRefPubMedGoogle Scholar
  16. 16.
    Novy JW (1997) Medicinal plants of the eastern region of Madagascar. J Ethnopharmacol 55:119–126CrossRefPubMedGoogle Scholar
  17. 17.
    Johns T, Mahunnah RL, Sanaya P, Chapman L, Ticktin T (1999) Saponins and phenolic content in plant dietary additives of a traditional subsistence community, the Batemi of Ngorongoro District, Tanzania. J Ethnopharmacol 66:1–10CrossRefPubMedGoogle Scholar
  18. 18.
    David CH, Michael ITN, Nigel JB, David RF (2001) The first total synthesis of toddaquinoline, an alkaloid from Toddalia asiatica. Tetrahedron 57:4447–4454CrossRefGoogle Scholar
  19. 19.
    Mukherjee AK, Basu S, Sarkar N, Ghosh AC (2001) Advances in cancer therapy with plant based natural products. Curr Med Chem 8:1467–1486PubMedGoogle Scholar
  20. 20.
    Bendixen C, Thomsen B, Alsner J, Westergaard O (1990) Camptothecin-stabilized topoisomerase I-DNA adducts cause premature termination of transcription. Biochemistry 29:5613–5619CrossRefPubMedGoogle Scholar
  21. 21.
    Diaz JF, Strobe R, Engelborghs Y, Souto AA, Andreu JM (2000) Molecular recognition of taxol by microtubules. Kinetics and thermodynamics of binding of fluorescent taxol derivatives to an exposed site. J Biol Chem 275:26265–26276CrossRefPubMedGoogle Scholar
  22. 22.
    Hanauske AR, Depenbrock H, Shirvani D, Rastetter J (1994) Effects of the microtubule-disturbing agents docetaxel (Taxotere), vinblastine and vincristine on epidermal growth factor-receptor binding of human breast cancer cell lines in vitro. Eur J Cancer 30A:1688–1694CrossRefPubMedGoogle Scholar
  23. 23.
    Tanaka E, Ho T, Kirschner MW (1995) The role of microtubule dynamics in growth cone motility and axonal growth. J Cell Biol 128:139–155CrossRefPubMedGoogle Scholar
  24. 24.
    Holden JA, Wall ME, Wani MC, Manikumar G (1999) Human DNA topoisomerase I: quantitative analysis of the effects of camptothecin analogs and the benzophenanthridine alkaloids nitidine and 6-ethoxydihydronitidine on DNA topoisomerase I-induced DNA strand breakage. Arch Biochem Biophys 370:66–76CrossRefPubMedGoogle Scholar
  25. 25.
    Wang LK, Johnson RK, Hecht SM (1993) Inhibition of topoisomerase I function by nitidine and fagaronine. Chem Res Toxicol 6:813–818CrossRefPubMedGoogle Scholar
  26. 26.
    Del Poeta M, Chen SF, Von HoffD, Dykstra CC, Wani MC, Manikumar G, Heitman J, Wall ME, Perfect JR (1999) Comparison of in vitro activities of camptothecin and nitidine derivatives against fungal and cancer cells. Antimicrob Agents Chemother 43:2862–2868PubMedGoogle Scholar
  27. 27.
    Hsiang YH, Liu LF (1988) Identification of mammalian DNA topoisomerase I as an intracellular target of the anticancer drug camptothecin. Cancer Res 48:1722–1726PubMedGoogle Scholar
  28. 28.
    Tung HT (2001) Analytical approaches for traditional Chinese medicines exhibiting antineoplastic activity. J Chromatogr 764:27–48CrossRefGoogle Scholar
  29. 29.
    McGuire WP, Hoskins WJ, Brady MF, Kucera PR, Partridge EE, Look KY, Clarke-Pearson DL, Davidson M (1996) Cyclophosphamide and cisplatin compared with paclitaxel and cisplatin in patients with stage III and stage IV ovarian cancer. N Engl J Med 334:1–6CrossRefPubMedGoogle Scholar
  30. 30.
    Flatters SJ, Bennett GJ (2004) Ethosuximide reverses paclitaxel- and vincristine-induced painful peripheral neuropathy. Pain 109:150–161CrossRefPubMedGoogle Scholar
  31. 31.
    Sheaff RJ, Groudine M, Gordon M, Roberts JM, Clurman BE (1997) Cyclin E-CDK2 is a regulator of p27Kip1. Genes Dev 11:1464–1478PubMedCrossRefGoogle Scholar
  32. 32.
    Lee SH, Hurwitz J (1990) Mechanism of elongation of primed DNA by DNA polymerase delta, proliferating cell nuclear antigen, and activator 1. Proc Natl Acad Sci USA 87:5672–5676PubMedCrossRefGoogle Scholar
  33. 33.
    Bennett M, Macdonald K, Chan SW, Luzio JP, Simari R, Weissberg P (1998) Cell surface trafficking of Fas: a rapid mechanism of p53-mediated apoptosis. Science 282:290–293CrossRefPubMedGoogle Scholar
  34. 34.
    Hill LL, Ouhtit A, Loughlin SM, Kripke ML, Ananthaswamy HN, Owen-Schaub LB (1999) Fas ligand: a sensor for DNA damage critical in skin cancer etiology. Science 285:898–900CrossRefPubMedGoogle Scholar
  35. 35.
    Fujita T, Maruyama M, Araya J, Sassa K, Kawagishi Y, Hayashi R, Matsui S, Kashii T, Yamashita N, Sugiyama E, Kobayashi M (2002) Hydrogen peroxide induces upregulation of Fas in human airway epithelial cells via the activation of PARP-p53 pathway. Am J Respir Cell Mol Biol 27:542–552PubMedGoogle Scholar
  36. 36.
    Miyashita T, Reed JC (1995) Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80:293–299CrossRefPubMedGoogle Scholar
  37. 37.
    Goldsmith ME, Gudas JM, Schneider E, Cowan KH (1995) Wild type p53 stimulates expression from the human multidrug resistance promoter in a p53-negative cell line. J Biol Chem 270:1894–1898CrossRefPubMedGoogle Scholar
  38. 38.
    Weber JD, Taylor LJ, Roussel MF, Sherr CJ, Bar-Sagi D (1999) Nucleolar Arf sequesters Mdm2 and activates p53. Nat Cell Biol 1:20–26CrossRefPubMedGoogle Scholar
  39. 39.
    Wu X, Bayle JH, Olson D, Levine AJ (1993) The p53-mdm-2 autoregulatory feedback loop. Genes Dev 7:1126–1132PubMedCrossRefGoogle Scholar
  40. 40.
    Kubbutat MH, Jones SN, Vousden KH (1997) Regulation of p53 stability by Mdm2. Nature 387:299–303CrossRefPubMedGoogle Scholar
  41. 41.
    Lohrum MA, Ashcroft M, Kubbutat MH, Vousden KH (2000) Contribution of two independent Mdm2-binding domains in p14(ARF) to p53 stabilization. Curr Biol 10:539–542CrossRefPubMedGoogle Scholar
  42. 42.
    Honda R, Yasuda H (1999) Association of p19(ARF) with Mdm2 inhibits ubiquitin ligase activity of Mdm2 for tumor suppressor p53. EMBO J 18:22–27CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Hironori Iwasaki
    • 1
  • Hirosuke Oku
    • 1
  • Ryo Takara
    • 1
  • Hanako Miyahira
    • 1
  • Kaoru Hanashiro
    • 2
  • Yasuhiko Yoshida
    • 2
  • Yasuhiro Kamada
    • 3
  • Tetsuya Toyokawa
    • 3
  • Kensaku Takara
    • 4
  • Masashi Inafuku
    • 1
  1. 1.Division of Molecular Biotechnology, Center of Molecular BioscienceUniversity of the RyukyusOkinawaJapan
  2. 2.Tropical Techno Center Limited OkinawaJapan
  3. 3.Okinawa Industrial Technology Center OkinawaJapan
  4. 4.Laboratory of Applied Biochemistry, Department of Bioscience and Biotechnology, Faculty of AgricultureUniversity of the RyukyusOkinawaJapan

Personalised recommendations