Kaempferol Induces Cell Cycle Arrest in HT-29 Human Colon Cancer Cells

BACKGROUND A greater intake of vegetables and fruits has been linked to a reduced incidence of colon cancer. Flavonoids are polyphenolic compounds which are broadly distributed in fruits and vegetables and display a remarkable spectrum of physiological activities, including anti-carcinogenic effects. The objective of this study was to determine the mechanisms by which kaempferol, a flavonol present in tea, apples, strawberries, and beans, inhibits the growth of HT-29 human colon cancer cells. METHODS To examine the effects of kaempferol on cell cycle progression in HT-29 cells, cells were treated with various concentrations (0-60 μmol/L) of kaempferol. Cell proliferation and DNA synthesis were evaluated by MTT assay and [(3)H]thymidine incorporation assay, respectively. Fluorescence-activated cell sorting analyses were conducted to calculate cell cycle phase distribution. Western blot analyses and in vitro kinase assays were used to estimate the expression of proteins involved in the regulation of cell cycle progression and the activity of cyclin-dependent kinase (CDK)s, respectively. RESULTS Kaempferol decreased viable cell numbers and [(3)H]thymidine incorporation into DNA of HT-29 cells in a dose-dependent manner. Kaempferol induced G1 cell cycle arrest within 6 h and G2/M arrest at 12 h. Kaempferol inhibited the activity of CDK2 and CDK4 as well as the protein expression of CDK2, CDK4, cyclins D1, cyclin E, and cyclin A, and suppressed the phosphorylation of retinoblastoma protein. Additionally, kaempferol decreased the levels of Cdc25C, Cdc2, and cyclin B1 proteins, as well as the activity of Cdc2. CONCLUSIONS The present results indicate that kaempferol induces G1 and G2/M cell cycle arrest by inhibiting the activity of CDK2, CDK4, and Cdc2. The induction of cell cycle arrest may be one of the mechanisms by which kaempferol exerts anti-carcinogenic effects in colon cancer cells.