Mechanism of sulforaphane-induced cell cycle arrest and apoptosis in human colon cancer cells

Nutr Cancer. 2004;48(2):198-206. doi: 10.1207/s15327914nc4802_10.

Abstract

Sulforaphane (SFN) is a natural micronutrient found in cruciferous vegetables that has been shown to possess antitumoral properties in carcinogen-treated rats. In vitro, SFN regulates phase II enzymes, cell cycle, and apoptosis. In the present study, we investigated the relationship between SFN induction of apoptosis and cell cycle arrest in HT29 human colon carcinoma cells. In previously published data, a significant increase in the G2/M phase of the cell cycle has been observed in SFN-treated cells that was associated with increased cyclin B1 protein levels. In the present study, our results show that SFN induced p21 expression. Moreover, preincubation of HT29 cells with roscovitine, a specific cdc2 kinase inhibitor, blocked the G2/M phase accumulation of HT29 cells treated with SFN and abolished its apoptotic effect (22.2 +/- 4 of floating cells in SFN-treated cells vs. 6.55 +/- 2 in cells treated with both SFN and roscovitine). These results suggest that the cdc2 kinase could be a key target for SFN in the regulation of G2/M block and apoptosis. Moreover, in SFN-treated cells the retinoblastoma tumor suppressor protein (Rb) is highly phosphorylated. Inhibition of the cdc2 kinase by roscovitine did not change the phosphorylation status of Rb in SFN-treated cells, suggesting that this cyclin-dependent kinase may not be involved. In our study, we did not observe any significant change in the proteasomal activity between control and SFN-treated cells. Moreover, inhibition of proteasomal activity through the use of MG132 diminished SFN-induced HT29 cell death, suggesting that the apoptotic effect of SFN requires a functional proteasome-dependent degradation system. In summary, we have elucidated part of the mechanism of action of SFN in the concomitant regulation of intestinal cell growth and death.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anticarcinogenic Agents / pharmacology*
  • Apoptosis / drug effects*
  • CDC2 Protein Kinase / antagonists & inhibitors*
  • CDC2 Protein Kinase / metabolism*
  • Cell Cycle / drug effects*
  • Flow Cytometry
  • G2 Phase / drug effects
  • HT29 Cells
  • Humans
  • Isothiocyanates
  • Mitosis / drug effects
  • Phosphorylation
  • Purines / pharmacology
  • Roscovitine
  • Thiocyanates / pharmacology*

Substances

  • Anticarcinogenic Agents
  • Isothiocyanates
  • Purines
  • Thiocyanates
  • Roscovitine
  • CDC2 Protein Kinase
  • sulforafan