Download PDF
Planta Med 2005; 71(11): 1030-1035
DOI: 10.1055/s-2005-873127
Original Paper
Pharmacology
© Georg Thieme Verlag KG Stuttgart · New York

The Lipophilic Extract of Hypericum perforatum Exerts Significant Cytotoxic Activity Against T24 and NBT-II Urinary Bladder Tumor Cells

Dimitris Skalkos1 , Nikolaos E. Stavropoulos2 , Ioannis Tsimaris2 , Eleni Gioti3 , Constantine D. Stalikas3 , Unyime O. Nseyo4 , Elli Ioachim5 , Niki J. Agnantis5
  • 1Department of Material Sciences & Engineering, University of Ioannina, Ioannina, Greece
  • 2Urology Department, ‘G. Hatzikosta’ General Hospital, Ioannina, Greece
  • 3Laboratory of Analytical Chemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
  • 4Division of Urology, Medical College of Virginia, Commonwealth University, Richmond, Virginia, USA
  • 5Department of Pathology, Medical School, University of Ioannina, Ioannina, Greece
Further Information

Publication History

Received: November 18, 2004

Accepted: May 30, 2005

Publication Date:
17 October 2005 (online)

Also available at
    Permissions and Reprints

Abstract

Hypericum perforatum L. (St. John’s wort) is a medicinal plant used for many pathologies, especially for the treatment of mild to moderate depression. In the present study we have investigated the cytotoxic activity of the locally collected (Epirus region) Hypericum perforatum L. against cultured T24 and NBT-II bladder cancer cell lines. The lipophilic extract of the herb, prepared using petroleum ether, induced apoptosis displaying LC50 values at concentrations as low as 4 and 5 μg/mL. A fraction of this extract displayed 60 % cell growth inhibition at a concentration of 0.95 μg/mL. Evaluating the importance of various biologically active components of the extract, it was found that hypericins (hypericin, pseudohypericin, etc.) were identified only in the methanolic (lipophobic) extract of the herb, and not in the active lipophilic extract. In addition, hyperforin concentrations in the lipophilic extract and its most active fraction, were 0.94 μg/mL, and 0.17 μg/mL, respectively, while the active cytotoxic concentration of pure hyperforin appeared in the range of 1.8 μg/mL - 5.0 μg/mL. Therefore, pure hyperforin does not seem to contribute significantly to the cytotoxicity activity. Chlorophylls were identified in low, not significantly different, concentrations in all extracts and fractions and were not correlated to the biological activity.

Owing to the combination of significant cytotoxic activity, natural abundance and low toxicity, the lipophilic extract of Hypericum perforatum holds the promise of being an interesting, new, antiproliferative agent against bladder cancer that deserves further investigation.

Abbreviations

HP:Hypericum perforatum L.

AS:aqueous solution

MS:methanolic solution

ME:methanolic extract

NPMF:non polar methanolic fraction

PMF:polar methanolic fraction

PEE:petroleum ether (lipophilic) extract

PEE (1,2,3,4,5,6,7):petroleum ether extract's fractions

PBS:phosphate buffered saline

DMSO:dimethyl sulfoxide

Key Words

Hypericum perforatum L. - St. John’s wort - hypericin - pseudohypericin - hyperforin - chlorophylls - bladder tumor

References

  • 1 Landis S H, Murray T, Bolden S, Wingo P A. Cancer Statistics, 1998.  CA Cancer J Clin. 1998;  48 6-29
    CrossrefPubMedSearch in Google Scholar
  • 2 Koss L G. Tumors of bladder. In: Koss LG, editor Diagnostic Cytology of the Urinary Tract With Histopathology and Clinical Correlations. New York; Lippincott-Raven 1995: pp 71-80
    Search in Google Scholar
  • 3 Kurth K H, Boufoux C, Sylvester R, van der Meijden A PM, Oosterlinck W, Brausi M. Treatment of superficial bladder tumors: achievements and needs.  Eur Urol. 2000;  37 1-9
    PubMedSearch in Google Scholar
  • 4 Nseyo U O, Lamn D L. Therapy of superficial bladder cancer.  Semin Oncol. 1996;  23 598-604
    PubMedSearch in Google Scholar
  • 5 Stavropoulos N E, Hastazeris K, Filiadis I, Michailidis I, Ioachim E, Liamis Z. et al . Intravesical instillations of interferon gamma in the prophylaxis of high risk superficial bladder cancer - results of a controlled prospective study.  Scand J Urol Nephrol. 2002;  36 218-22
    PubMedSearch in Google Scholar
  • 6 Prout GR J r, Barton B A, Grifin P P, Friedell G H. Treated history of non-invasive grade 1 transitional cell carcinoma.  J Urol. 1992;  148 1413-9
    PubMedSearch in Google Scholar
  • 7 Nseyo U O, DeHaven J, Dougherty T J, Potter W R, Merill D L, Lundahl S L. et al . Photodynamic therapy (PDT) in the treatment of patients with resistant superficial bladder cancer: a long-term experience.  J Clin Laser Med Surg. 1998;  16 61-8
    PubMedSearch in Google Scholar
  • 8 Blumenthal M, Busse W R, Goldberg A, Gruenwald J, Hall T, Riggins C W, Rister R S, editors . The complete German Commission E monographs: therapeutic guide to herbal medicines. Boston; American Botanical Council 1998
    Search in Google Scholar
  • 9 St John’s W ort. (Hypericum perforatum). In: Upton R editor American herbal pharmacopoeia (AHP). St. Cruz; 1997: pp 1-32
    Search in Google Scholar
  • 10 Malamas M, Marselos M. The tradition of medicinal plants in Zagori, Epirus (Northwestern Greece).  J Ethnopharmacol. 1992;  37 197-203
    CrossrefPubMedSearch in Google Scholar
  • 11 Brantner A, Della Loggia R, Sosa S, Kartnig T. Untersuchungen zur antiphlogistichen Wirkung von Hypericum perforatum L.  Sci Pharm. 1994;  62 7-8
    PubMedSearch in Google Scholar
  • 12 Shakirova K K, Garagulya A D, Khazanovich R L. Antimicrobial properties of some species of St. John’s wort cultivated in Uzbekistan.  Mikrobiol Zh. 1970;  32 494-7
    PubMedSearch in Google Scholar
  • 13 Yip L, Hudson J B, Gruszecka-Kowalik , Zalkow L H, Towers G H. Antiviral activity of a derivative of the photosensitive compound hypericin.  Phytomedicine III. 1996;  2 185-90
    PubMedSearch in Google Scholar
  • 14 Harrer G, Sommer H. Treatment of mild/moderate depressions with Hypericum .  Phytomedicine. 1994;  1 3-8
    PubMedSearch in Google Scholar
  • 15 Linde K, Murlow C J, Berner M, Egger M. St John’s wort for depression.  Cochrane Database Syst Rev. 2005;  18 CD000448
    PubMedSearch in Google Scholar
  • 16 Bilia A R, Gallori S, Vincieri F F. St John’s wort and depression efficacy, safety and tolerability - an update.  Life Sci. 2002;  70 3077-96
    CrossrefPubMedSearch in Google Scholar
  • 17 Pietta P, Gardana C, Pietta A. Comparative evaluation of St. John’s wort from different Italian regions.  Il Farmaco. 2001;  56 491-6
    CrossrefPubMedSearch in Google Scholar
  • 18 Singer A, Wonneman M, Mueller W E. Hyperforin, a major antidepressant constituent of St. John’s wort inhibits serotonin uptake by elevating free intracellular Na+ 1 .  J Pharmacol Exp Ther. 1999;  290 1363-8
    PubMedSearch in Google Scholar
  • 19 Butterweck V, Petereit F, Winterhoff H, Nahrstedt A. Solubilized-hypericin and pseudohypericin from Hypericum perforatum exert anti-depressant activity in the forced swimming test.  Planta Med. 1998;  64 291-4
    Thieme ConnectPubMedSearch in Google Scholar
  • 20 Butterweck V, Jurgenliemk G, Nahrstedt A, Winterhoff H. Flavonoids from Hypericum perforatum show antidepressant activity in the forced swimming test.  Planta Med. 2000;  66 3-6
    Thieme ConnectPubMedSearch in Google Scholar
  • 21 Skalkos D, Tatsis E, Gerothanasis I P, Troganis A. Towards a consensus structure of hypericin in solution: direct evidence for a single tautomer and different ionization states in protic and nonprotic solvents by the use of variable temperature gradient 1H NMR.  Tetrahedron. 2002;  58 4925-9
    CrossrefPubMedSearch in Google Scholar
  • 22 United States P harmacopoeia. St. John’s wort monograph, 9th Supplement. 1998: pp 4709-4710
    Search in Google Scholar
  • 23 AOAC method 9 42.04, Chlorophylls in p lants. In: Cunniff P, editor Official Methods of Analysis of AOAC International. Gaithersburg, MD; 1998
    Search in Google Scholar
  • 24 Fotsis T, Pepper M S, Aktas E, Breit S, Rasku S, Adlercreutz H. et al . Flavonoids, dietary-derived inhibitors of cell proliferation and in vitro angiogenesis.  Cancer Res. 1997;  57 2916-21
    PubMedSearch in Google Scholar
  • 25 Hostanska K, Reichling J, Bommer S, Weber M, Saller R. Aqueous ethanolic extract of St. John’s wort (Hypericum perforatum L.) induces growth inhibition and apoptosis in human malignant cells in vitro .  Pharmazie. 2002;  57 323-31
    PubMedSearch in Google Scholar
  • 26 Hostanska K, Reichling J, Bommer S, Weber M, Saller R. Hyperforin a constituent of St John’s wort (Hypericum perforatum L.) extract induces apoptosis by triggering activation of caspases and with hypericin synergistically exerts cytotoxicity towards human malignant cell lines.  Eur J Pharm Biopharm. 2003;  56 121-32
    CrossrefPubMedSearch in Google Scholar
  • 27 Schempp C M, Kirkin V, Simon-Haarhaus B, Kersten A, Kiss J, Termeer C C. et al . Inhibition of tumour cell growth by hyperforin, a novel anticancer drug from St. John’s wort that acts by induction of apoptosis.  Oncogene. 2002;  21 1242-5
    CrossrefPubMedSearch in Google Scholar

Dimitris Skalkos

Department of Material Sciences & Engineering

University of Ioannina

Ioannina GR-45110

Greece

Phone: +32-651-097-262

Fax: +32-651-097-056

Email: dskalkos@cc.uoi.gr

    www.thieme-connect.de/ejournals/toc/plantamedica