• Open Access

Mechanism of inhibition of tumor angiogenesis by β-hydroxyisovalerylshikonin

Authors

  • Yusuke Komi,

    1. Molecular Ligand Biology Research Team, Chemical Genomics Research Group, Chemical Biology Department, RIKEN Advanced Science Institute, 2-1 Hiroshima, Wako, Saitama 351-0198;
    2. Vascular Medicine and Geriatrics, Tokyo Medical and Dental University Graduate School, 1-5-45 Yushima, Bunkyo, Tokyo 113-8519;
    Search for more papers by this author
  • Yasuhiro Suzuki,

    1. Molecular Ligand Biology Research Team, Chemical Genomics Research Group, Chemical Biology Department, RIKEN Advanced Science Institute, 2-1 Hiroshima, Wako, Saitama 351-0198;
    Search for more papers by this author
    • 11

      Present address: Department of Vascular Biology, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryo-machi, Aoba, Sendai, Miyagi 980-8575, Japan

  • Mariko Shimamura,

    1. Medical R & D Center, Tokyo Metropolitan Institute of Medical Science, 3-18-22 Honkomagome, Bunkyo, Tokyo 113-8613;
    Search for more papers by this author
    • 12

      Present address: Tokyo Metropolitan Institute of Public Health, Tokyo Metropolitan Infectious Disease Surveillance Center, 3-24-1 Hyakunincho, Shinjuku, Tokyo 169-0073, Japan

  • Sachiko Kajimoto,

    1. Laboratory of Biological Chemistry, School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa, Tokyo 142-8555;
    Search for more papers by this author
  • Shigeo Nakajo,

    1. Laboratory of Biochemistry, Yokohama College of Pharmacy, 601 Minato, Totsuka, Yokohama, Kanagawa 245-0066;
    Search for more papers by this author
  • Michitaka Masuda,

    1. Department of Structural Analysis, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565;
    Search for more papers by this author
  • Masabumi Shibuya,

    1. Department of Genetics, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato, Tokyo 108-8639, Japan;
    Search for more papers by this author
    • 13

      Present address: Department of Molecular Oncology, Tokyo Medical and Dental University Graduate School, 1-5-45 Yushima, Bunkyo, Tokyo 113-8519, Japan

  • Hiroyuki Itabe,

    1. Laboratory of Biological Chemistry, School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa, Tokyo 142-8555;
    Search for more papers by this author
  • Kentaro Shimokado,

    1. Vascular Medicine and Geriatrics, Tokyo Medical and Dental University Graduate School, 1-5-45 Yushima, Bunkyo, Tokyo 113-8519;
    Search for more papers by this author
  • Peter Oettgen,

    1. Division of Cardiology, Beth Israel Deaconess Medical Center, 330 Brookline Ave., Boston, MA 02115, USA;
    Search for more papers by this author
  • Kazuyasu Nakaya,

    1. Animal Cell Biological Engineering, Niigata University of Pharmacy and Applied Life Science, Niitsu, 265-1 Higashijima, Niigata 956-8603, Japan
    Search for more papers by this author
  • Soichi Kojima

    Corresponding author
    1. Molecular Ligand Biology Research Team, Chemical Genomics Research Group, Chemical Biology Department, RIKEN Advanced Science Institute, 2-1 Hiroshima, Wako, Saitama 351-0198;
    Search for more papers by this author

To whom correspondence should be addressed. E-mail: skojima@postman.riken.go.jp

Abstract

Shikonin and β-hydroxyisovalerylshikonin (β-HIVS) from Lithospermum erythrorhizon inhibit angiogenesis via inhibition of vascular endothelial growth factor receptors (VEGFR) in an adenosine triphosphate-non-competitive manner, although the underlying molecular mechanism has not been fully understood. In the present study, we found that β-HIVS inhibited angiogenesis within chicken chorioallantoic membrane approximately threefold more efficiently than shikonin. β-HIVS also significantly inhibited angiogenesis in two other assays, induced either by Lewis lung carcinoma cells implanted in mouse dorsal skin or by VEGF in s.c. implanted Matrigel plugs and metastasis of Lewis lung carcinoma cells to lung. Therefore, using β-HIVS as a bioprobe, we investigated the molecular mechanism of shikonin's anti-angiogenic actions. β-HIVS inhibited the phosphorylation and expression of VEGFR2 and Tie2 without affecting VEGFR1 and fibroblast growth factor receptor 1 levels. β-HIVS suppressed the phosphorylation but not the expression of extracellular signal-regulated kinase, and an Sp1-dependent transactivation of the VEGFR2 and Tie2 promoters, thereby suppressing the proliferation of vascular endothelial and progenitor cells. This was mimicked by an Sp1 inhibitor mithramycin A and partially rescued by Sp1 overexpression. These results implicate potential use of shikonin and β-HIVS as leading compounds for clinical application in the future by virtue of their unique properties including: (i) inhibition of VEGFR2 and Tie2 phosphorylation in an adenosine triphosphate-non-competitive manner; (ii) simultaneous inhibition of the phosphorylation and expression of VEGFR2 and Tie2; and (iii) bifunctional inhibition of the growth in endothelial cells and vascular remodeling. (Cancer Sci 2009; 100: 269–277)

Ancillary