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

Abstract

We have shown previously that diallyl trisulfide (DATS), a constituent of processed garlic, inhibits proliferation of PC-3 and DU145 human prostate cancer cells by causing G(2)-M phase cell cycle arrest in association with inhibition of cyclin-dependent kinase 1 activity and hyperphosphorylation of Cdc25C at Ser(216). Here, we report that DATS-treated PC-3 and DU145 cells are also arrested in mitosis as judged by microscopy following staining with anti-alpha-tubulin antibody and 4',6-diamidino-2-phenylindole and flow cytometric analysis of Ser(10) phosphorylation of histone H3. The DATS treatment caused activation of checkpoint kinase 1 and checkpoint kinase 2, which are intermediaries of DNA damage checkpoints and implicated in Ser(216) phosphorylation of Cdc25C. The diallyl trisulfide-induced Ser(216) phosphorylation of Cdc25C as well as mitotic arrest were significantly attenuated by knockdown of check-point kinase 1 protein in both PC-3 and DU145 cells. On the other hand, depletion of checkpoint kinase 2 protein did not have any appreciable effect on G(2) or M phase arrest or Cdc25C phosphorylation caused by diallyl trisulfide. The lack of a role of checkpoint kinase 2 in diallyl trisulfide-induced phosphorylation of Cdc25C or G(2)-M phase cell cycle arrest was confirmed using HCT-15 cells stably transfected with phosphorylation-deficient mutant (T68A mutant) of checkpoint kinase 2. In conclusion, the results of the present study suggest existence of a checkpoint kinase 1-dependent mechanism for diallyl trisulfide-induced mitotic arrest in human prostate cancer cells.

• Authors: Anna Herman-Antosiewicz, Shivendra V Singh
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 280 Issue 31 Pg. 28519-28 (Aug 05 2005) ISSN: 0021-9258 [Print] United States
• PMID: 15961392 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Allyl Compounds
• RNA, Small Interfering
• Recombinant Proteins
• Sulfides
• diallyl trisulfide
• Protein Kinases
• CHEK1 protein, human
• Checkpoint Kinase 1

Topics

• Allyl Compounds (pharmacology)
• Cell Cycle (drug effects, physiology)
• Cell Division (drug effects)
• Cell Line, Tumor
• Checkpoint Kinase 1
• Humans
• Male
• Mitosis (drug effects, physiology)
• Prostatic Neoplasms
• Protein Kinases (genetics, metabolism)
• RNA, Small Interfering (genetics)
• Recombinant Proteins (metabolism)
• Sulfides (pharmacology)
• Transfection