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Checkpoint kinase 1 regulates diallyl trisulfide-induced mitotic arrest in human prostate cancer cells

Anna Herman-Antosiewicz1, Shivendra V Singh

  • 1Department of Pharmacology and University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.

The Journal of Biological Chemistry
|June 18, 2005
PubMed

Insights

Diallyl trisulfide (DATS), found in garlic, halts prostate cancer cell division by triggering mitotic arrest. This process relies on checkpoint kinase 1, not checkpoint kinase 2, highlighting a specific pathway for DATS

Area of Science:

  • Oncology
  • Cell Biology
  • Biochemistry

Background:

  • Diallyl trisulfide (DATS), a garlic compound, previously demonstrated inhibition of prostate cancer cell proliferation.
  • DATS induced G(2)-M phase cell cycle arrest by inhibiting cyclin-dependent kinase 1 and phosphorylating Cdc25C at Ser(216).
  • The precise molecular mechanisms underlying DATS-induced mitotic arrest require further elucidation.

Purpose of the Study:

  • To investigate the role of checkpoint kinase 1 (Chk1) and checkpoint kinase 2 (Chk2) in DATS-induced mitotic arrest in human prostate cancer cells.
  • To determine the specific pathway mediating DATS' effects on cell cycle regulation and Cdc25C phosphorylation.

Main Methods:

  • Microscopy with anti-alpha-tubulin antibody and 4',6-diamidino-2-phenylindole staining to assess mitotic arrest.
  • Flow cytometry to analyze histone H3 phosphorylation at Ser(10).
  • Western blotting and siRNA-mediated knockdown of Chk1 and Chk2 in PC-3, DU145, and HCT-15 cells.

Main Results:

  • DATS treatment induced clear mitotic arrest in PC-3 and DU145 cells, confirmed by microscopy and histone H3 phosphorylation.
  • DATS activated both Chk1 and Chk2, key intermediaries in DNA damage checkpoints.
  • Chk1 knockdown significantly attenuated DATS-induced Cdc25C phosphorylation at Ser(216) and mitotic arrest, while Chk2 depletion had no significant effect.

Conclusions:

  • The study identifies a Chk1-dependent mechanism responsible for DATS-induced mitotic arrest in human prostate cancer cells.
  • Chk1 plays a critical role in mediating the phosphorylation of Cdc25C at Ser(216) and subsequent G(2)-M phase arrest induced by DATS.
  • Chk2 is not essential for DATS-induced mitotic arrest or Cdc25C phosphorylation in these cancer cell models.

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