Planta Med 2006; 72(5): 424-429
DOI: 10.1055/s-2005-916259
Original Paper
Pharmacology
© Georg Thieme Verlag KG Stuttgart · New York

Induction of Apoptosis by Isoflavonoids from the Leaves of Millettia taiwaniana in Human Leukemia HL-60 Cells

Chihiro Ito1 , Tomiyasu Murata2 , Masataka Itoigawa1 , 3 , Keisuke Nakao1 , Minako Kumagai1 , Norio Kaneda2 , Hiroshi Furukawa1
  • 1Department of Medicinal Chemistry, Faculty of Pharmacy, Meijo University, Nagoya, Japan
  • 2Department of Analytical Neurobiology, Faculty of Pharmacy, Meijo University, Nagoya, Japan
  • 3Faculty of Human Wellness, Tokai Gakuen University, Nagoya, Japan
Further Information

Publication History

Received: March 23, 2005

Accepted: October 31, 2005

Publication Date:
17 February 2006 (online)

Abstract

We have isolated two new isoflavonoids, millewanin-F (1) and furowanin-A (2), together with five known isoflavonoids from the leaves of Millettia taiwaniana Hayata (Leguminosae) and examined their effects on the growth of human leukemia HL-60 cells. Among the isolated isoflavonoids, furowanin-A (2), warangalone (3), isoerysenegalensein-E (4), and euchrenone b10 (6) showed significant cytotoxicity against HL-60 cells. After treatment with three of the cytotoxic isoflavonoids, furowanin-A (2), warangalone (3), and isoerysenegalensein-E (4), fluorescence microscopy with Hoechst 33 342 staining revealed that the percentage of apoptotic cells with fragmented nuclei and condensed chromatin increased in a time-dependent manner. In addition, the activities of caspase-9 and caspase-3 were also enhanced in a time-dependent manner upon treatment with the isoflavonoids 2, 3, and 4. Caspase-9 and caspase-3 inhibitors suppressed apoptosis induced by isoflavonoids 2, 3, and 4. These results suggest that the isoflavonoids induced apoptosis in HL-60 cells through activation of the caspase-9/caspase-3 pathway, which is triggered by mitochondrial dysfunction.

References

  • 1 Konoshima T, Takasaki M, Kozuka M, Tokuda H, Nishino H, Matsuda E. et al . Anti-tumor promoting activities of isoflavonoids from Wistaria brachybotrys .  Biol Pharm Bull. 1997;  20 865-8
  • 2 Ito C, Itoigawa M, Tan H T, Tokuda H, Yang Mou X, Mukainaka T. et al . Anti-tumor-promoting effects of isoflavonoids on Epstein-Barr virus activation and two-stage mouse skin carcinogenesis.  Cancer Lett. 2000;  152 187-92
  • 3 Jung S H, Lee Y S, Lee S, Lim S S, Kim Y S, Ohuchi K. et al . Anti-angiogenic and anti-tumor activities of isoflavonoids from the rhizomes of Belamcanda chinensis .  Planta Med. 2003;  69 617-22
  • 4 Ito C, Itoigawa M, Kojima N, Tokuda H, Hirata T, Nishino H. et al . Chemical constituents of Millettia taiwaniana: structure elucidation of five new isoflavonoids and their cancer chemopreventive activity.  J Nat Prod. 2004;  67 1125-30
  • 5 Guo Q, Rimbach G, Moini H, Weber S, Packer L. ESR and cell culture studies on free radical-scavenging and antioxidant activities of isoflavonoids.  Toxicology. 2002;  179 171-80
  • 6 Laupattarakasem P, Houghton P J, Hoult J R. Anti-inflammatory isoflavonoids from the stems of Derris scandens .  Planta Med. 2004;  70 496-501
  • 7 Kumi-Diaka J, Saddler-Shawnette S, Aller A, Brown J. Potential mechanism of phytochemical-induced apoptosis in human prostate adenocarcinoma cells: therapeutic synergy in genistein and beta-lapachone combination treatment.  Cancer Cell Int. 2004;  4 5
  • 8 Fomum Z T, Ayafor J F, Wandji J. Erythrisenegalone, a prenylated-flavanone from Erythrina senegalensis .  Phytochemistry. 1985;  24 3075-6
  • 9 El-Masry S, Amer M E, Abdel-Kader M S, Zaatout H H. Prenylated flavonoids of Erythrina lysistemon grown in Egypt.  Phytochemistry. 2002;  60 783-7
  • 10 Tanaka H, Doi M, Etoh H, Watanabe N, Shimizu H, Hirata M. et al . Revised structures for senegalensin and euchrenone b10 .  J Nat Prod. 2001;  64 1336-40
  • 11 Singhal A K, Sharma R P, Thyagarajan G, Herz W, Govindan S V. New prenylated isoflavones and a prenylated dihydroflavonol from Millettia pachycarpa .  Phytochemistry. 1980;  19 929-34
  • 12 Patel T, Gores G J, Kaufmann S H. The role of proteases during apoptosis.  FASEB J. 1996;  10 587-97
  • 13 Wang X. The expanding role of mitochondria in apoptosis.  Genes Dev. 2001;  15 2922-33
  • 14 Yin X M. Signal transduction mediated by Bid, a pro-death Bcl-2 family proteins, connects the death receptor and mitochondria apoptosis pathways.  Cell Res. 2000;  10 161-7
  • 15 Gross A, McDonnell J M, Korsmeyer S J. BCL-2 family members and the mitochondria in apoptosis.  Genes Dev. 1999;  13 1899-911

Prof. Dr. Masataka Itoigawa

Tokai Gakuen University

2-901 Nakahira

Tempaku-ku

Nagoya 468-8514

Japan

Phone: +81-52-801-1201

Fax: +81-52-804-1044

Email: itoigawa@tokaigakuen-u.ac.jp

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