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Planta Med 2011; 77(16): 1788-1793
DOI: 10.1055/s-0030-1271157
Biological and Pharmacological Activity
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

Cytotoxicity of Ethanolic Extracts of Artemisia annua to Molt-4 Human Leukemia Cells

Narendra P. Singh1 , Jorge F. S. Ferreira2 , Ji Sun Park1 , Henry C. Lai1
  • 1Department of Bioengineering, University of Washington, Seattle, WA, USA
  • 2Appalachian Farming Systems Research Center, USDA-ARS, Beaver, WV, USA
Further Information

Publication History

received February 2, 2011 revised April 29, 2011

accepted May 5, 2011

Publication Date:
14 June 2011 (online)

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Abstract

Although dihydroartemisinin (DHA) and other artemisinin derivatives have selective toxicity towards cancer cells, Artemisia annua (A. annua) extracts containing artemisinin have not been evaluated for their anticancer potential. Our main goal was to assess the anticancer effect of ethanolic leaf extracts of A. annua from Brazilian and Chinese origins (with DHA as a comparison) on normal and cancer cells. Leukocytes and leukemia (Molt-4) cells were counted at 0, 24, 48, and 72 hr after treatment with extracts having artemisinin concentrations of 0, 3.48, 6.96, and 13.92 µg/mL. Also, we assessed the antioxidant capacity of these extracts using the oxygen radical absorbance capacity (ORAC) test. Both extracts had high antioxidant capacity and toxicity towards Molt-4 cells. DHA was significantly more potent (p < 0.05) in killing Molt-4 cells than Brazilian extract at 48 and 72 hr and Chinese extract at 72 hr. In Molt-4 cells, LD50 values for Brazilian and Chinese extracts were comparable at all time points and not significantly different from DHA at 24 hr. In leukocytes, DHA, Chinese extract, and Brazilian extract had LD50 values of 760.42, 13.79, and 28.23 µg/mL of artemisinin, respectively, indicating a better safety index for the Brazilian extract compared to that of the Chinese extract at 24 hr. However, at 48 and 72 hr, the toxicity in leukocytes for any of the treatment groups was not significantly different. These experiments suggest that these extracts may have potential application in cancer treatment.

Key words

Artemisia annua - Asteraceae - ethanolic extracts - antioxidant - flavonoids - synergism - leukemia - MOLT-4

References

  • 1 de Ridder S, van der Kooy F, Verpoorte R. Artemisia annua as a self-reliant treatment for malaria in developing countries.  J Ethnopharmacol. 2008;  120 302-314
    CrossrefPubMedGoogle Scholar
  • 2 Lai H, Singh N. Selective cancer cell cytotoxicity from exposure to dihydroartemisinin and holotransferrin.  Cancer Lett. 1995;  91 41-46
    CrossrefPubMedGoogle Scholar
  • 3 Singh N, Lai H. Selective toxicity of dihydroartemisinin and holotransferrin toward human breast cancer cells.  Life Sci. 2001;  70 49-56
    CrossrefPubMedGoogle Scholar
  • 4 Moore J C, Lai H, Li J R, Ren R L, McDougall J A, Singh N P, Chou C K. Oral administration of dihydroartemisinin and ferrous sulfate retarded implanted fibrosarcoma growth in the rat.  Cancer Lett. 1995;  98 83-87
    PubMedGoogle Scholar
  • 5 Disbrow G L, Baege A C, Kierpiec K A. Dihydroartemisinin is cytotoxic to papillomavirus-expressing epithelial cells in vitro and in vivo.  Cancer Res. 2005;  65 10854-10861
    CrossrefPubMedGoogle Scholar
  • 6 Lai H, Nakase I, Lacoste E, Singh N P, Sasaki T. Artemisinin-transferrin conjugate retards growth of breast tumors in the rat.  Anticancer Res. 2009;  29 3807-3810
    PubMedGoogle Scholar
  • 7 Hou J, Wang D, Zhang R, Wang H. Experimental therapy of hepatoma with artemisinin and its derivatives: in vitro and in vivo activity, chemosensitization, and mechanisms of action.  Clin Cancer Res. 2008;  14 5519-5530
    CrossrefPubMedGoogle Scholar
  • 8 Riganti C, Doublier S, Viarisio D, Miraglia E, Pescarmona G, Ghigo D, Bhosia A. Artemisinin induces doxorubicin resistance in human colon cancer cells via calcium-dependent activation of HIF-1alpha and P-glycoprotein overexpression.  Brit J Pharmacol. 2009;  156 1054-1066
    CrossrefPubMedGoogle Scholar
  • 9 Reungpatthanaphong P, Mankhetkorn S. Modulation of multidrug resistance by artemisinin, artesunate and dihydroartemisinin in K562/adr and GLC4/adr resistant cell lines.  Biol Pharm Bull. 2002;  25 1555-1561
    CrossrefPubMedGoogle Scholar
  • 10 Perazzo F F, Lima L M, Maistro E L, Carvalho J E, Rehder V L G, Carvalho J C T. Effect of Artemisia annua L. leaves essential oil and ethanol extract on behavioral assays.  Braz J Pharmacognosy. 2008;  18 686-689
    PubMedGoogle Scholar
  • 11 Bilia A R, Melillo de Malgalhaes P, Bergonzi M C, Vincieri F F. Simultaneous analysis of artemisinin and flavonoids of several extracts of Artemisia annua L. obtained from a commercial sample and a selected cultivar.  Phytomedicine. 2006;  13 487-493
    CrossrefPubMedGoogle Scholar
  • 12 Ferreira J F S. Artemisia species in small ruminant production: their potential antioxidant and anthelmintic effects. Appalachian Workshop and Research Update: Improving Small Ruminant Grazing Practices Beaver, WV; 2009
    Google Scholar
  • 13 Walle T. Methoxylated flavones, a superior cancer chemopreventive flavonoid subclass?.  Semin Cancer Biol. 2007;  17 354-362
    CrossrefPubMedGoogle Scholar
  • 14 Räth K, Taxis K, Walz G, Gleiter C H, Li S M, Heide L. Pharmacokinetic study of artemisinin after oral intake of a traditional preparation of Artemisia annua L. (Annual Wormwood).  Am J Trop Med Hyg. 2004;  70 128-132
    PubMedGoogle Scholar
  • 15 Ferreira J F S. Seasonal and post-harvest accumulation of artemisinin, artemisinic acid, and dihydroartemisinic acid in three accessions of Artemisia annua cultivated in West Virginia, USA.  Planta Med. 2008;  74 310-311
    PubMedGoogle Scholar
  • 16 Prior R L, Wu X L, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements.  J Agric Food Chem. 2005;  53 4290-4302
    CrossrefPubMedGoogle Scholar
  • 17 Ferreira J F S, Gonzalez J M. Analysis of underivatized artemisinin and related sesquiterpene lactones by high-performance liquid chromatography with ultraviolet detection.  Phytochem Anal. 2008;  20 91-97
    PubMedGoogle Scholar
  • 18 Wallaart T E, Pras N, Beekman A C, Quax W J. Seasonal variation of artemisinin and its biosynthetic precursors in plants of Artemisia annua of different geographical origin: proof for the existence of chemotypes.  Planta Med. 2000;  66 57-62
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Squires J M, Ferreira J F S, Lindsay D S, Zajac A M. Effects of artemisinin and Artemisia and Artemisia extracts on Haemonchus contortus in gerbils (Meriones unguiculatus).  Vet Parisitol. 2011;  175 103-108
    PubMedGoogle Scholar
  • 20 Wang J, Guo L L, Wang Y Y. Study on combination components and effectiveness of Chinese traditional herbal formulas.  Zhongguo Zhong Yao Za Zhi. 2006;  31 5-9
    PubMedGoogle Scholar
  • 21 Lee K H. Research and future trends in the pharmaceutical development of medicinal herbs from Chinese medicine.  Public Health Nutr. 2000;  3 515-522
    PubMedGoogle Scholar
  • 22 Olsson M E, Gustavsson K, Andersson S, Nilsson Å, Duan R. Inhibition of cancer cell proliferation in vitro by fruit and berry extracts and correlations with antioxidant levels.  J Agric Food Chem. 2004;  52 7264-7271
    CrossrefPubMedGoogle Scholar
  • 23 Ferreira J F S, Luthria D L, Sasaki T, Heyerick A. Flavonoids from Artemisia annua L. as antioxidants and their potential synergism with artemisinin against malaria and cancer.  Molecules. 2010;  15 3135-3170
    CrossrefPubMedGoogle Scholar
  • 24 Zheng G Q. Cytotoxic terpenoids and flavonoids from Artemisia annua.  Planta Med. 1994;  60 54-57
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Amin A, Buratovich M. The anti-cancer charm of flavonoids: a cup-of-tea will do!.  Anticancer Drug Discov. 2007;  2 109-117
    PubMedGoogle Scholar
  • 26 Svensson U S H, Ashton M. Identification of the human cytochrome P450 enzymes involved in the in vitro metabolism of artemisinin.  Br J Clin Pharmacol. 1999;  48 528-535
    PubMedGoogle Scholar
  • 27 Aldieri E, Atragene D, Bergandi L, Riganti C, Costamagna C, Bosia A, Ghigo D. Artemisinin inhibits inducible nitric oxide synthase and nuclear factor NF-κB activation.  FEBS Lett. 2003;  552 141-144
    CrossrefPubMedGoogle Scholar
  • 28 Ulrich-Merzenich G, Panek D, Zeitler H, Wagner H, Vetter H. New perspectives for synergy research with the “omic”-technologies.  Phytomedicine. 2009;  16 495-508
    CrossrefPubMedGoogle Scholar
  • 29 Willcox M L, Bodeker G. Plant-based malaria control: research initiative on traditional antimalarial methods.  Parasitol Today. 2000;  16 220-221
    CrossrefPubMedGoogle Scholar

Dr. Narendra P. Singh

Department of Bioengineering
University of Washington

Box 355061

Seattle, WA 98195-5061

USA

Phone: +1 206 68 5 20 60

Fax: +1 206 68 5 39 25

Email: narendra@u.washington.edu