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Cancer Chemotherapy and Pharmacology

, Volume 51, Issue 3, pp 202–208 | Cite as

Ritterazine B, a new cytotoxic natural compound, induces apoptosis in cancer cells

  • Takefumi Komiya
  • Nobuhiro Fusetani
  • Shigeki Matsunaga
  • Akihito Kubo
  • Frederic J. Kaye
  • Michael J. Kelley
  • Kenji Tamura
  • Makoto Yoshida
  • Masahiro Fukuoka
  • Kazuhiko Nakagawa
Original Article
  • 229 Downloads

Abstract

Purpose

Ritterazine B, one of the ritterazine analogues extracted from Ritterella tokioka, has been shown to be chemically similar to cephalostatin 1, and among the ritterazine derivatives is the most cytotoxic to P388 murine leukemia cells. The objective of this study was to determine the cytotoxicity of ritterazine B to non-small-cell lung cancer (NSCLC) cells in vitro and its effects on the cell cycle and apoptosis.

Methods

The cytotoxicity of ritterazine B against PC14 NSCLC cells was investigated using a 4-day MTT assay. Morphological changes in cells after exposure to this compound were evaluated by phase-contrast microscopy. The effects on the cell cycle of HL-60 leukemia cells and PC14 cells were elucidated by flow cytometry and an in vitro CDK/cyclin kinase assay. Induction of apoptosis in HL-60 cells was assessed using the TUNEL assay and Hoechst 33342 staining. In addition, molecules involved in apoptosis were evaluated by Western blotting.

Results

Ritterazine B exerted strong cytotoxic effects against PC14 cells with a mean GI50 of 75.1 nM. Cell cycle analysis showed that ritterazine B caused accumulation of HL-60 and PC14 cells at the G2/M checkpoint. Furthermore, ritterazine B-treated HL-60 cells became multinucleated, and at a concentration of 20 nM this resulted in the onset of apoptosis. Neither cleavage of caspase target molecules nor phosphorylation of bcl-2 were observed in ritterazine B-treated HL-60 cells.

Conclusions

These results indicate that ritterazine B might be a potent inducer of apoptosis acting via a novel antimitotic mechanism.

Keywords

Ritterazine Cell cycle arrest Apoptosis Caspase 

Abbreviations

CDK

Cyclin-dependent kinase

DMSO

Dimethylsulfoxide

ECL

Enhanced chemiluminescence

FITC

Fluorescein isothiocyanate

GI50

50% growth inhibition

IC50

50% enzyme inhibition

MTT

3-(4,5-Dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide

NSCLC

Non-small-cell lung cancer

PARP

Poly(ADP-ribose) polymerase

PBS

Phosphate-buffered saline

PI

Propidium iodide

TUNEL

Terminal deoxynucleotidyl nick end-labeling

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Takefumi Komiya
    • 1
    • 3
  • Nobuhiro Fusetani
    • 2
  • Shigeki Matsunaga
    • 2
  • Akihito Kubo
    • 3
  • Frederic J. Kaye
    • 3
  • Michael J. Kelley
    • 4
  • Kenji Tamura
    • 1
  • Makoto Yoshida
    • 1
  • Masahiro Fukuoka
    • 1
  • Kazuhiko Nakagawa
    • 1
  1. 1.Division of Medical OncologyKinki University School of MedicineOsakasayama CityJapan
  2. 2.Laboratory of Marine BiochemistryThe University of TokyoTokyoJapan
  3. 3.Genetics Branch, Center for Cancer Research, National Naval Medical CenterNational Cancer InstituteBethesdaUSA
  4. 4.Department of MedicineDuke University School of MedicineDurhamUSA

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