Kaempferol induced inhibition of HL-60 cell growth results from a heterogeneous response, dominated by cell cycle alterations

Dietary flavonoids may be exploitable as chemotherapeutics and preventatives for critical health conditions, including cancer. Antiproliferative effects are commonly ascribed to such compounds but ambiguity exists as to the principal mechanism of action and the universal benefit of exposure, particularly at high concentrations. Here, we identify heterogeneous responses within HL-60 promyelocytic leukaemia cells that explain contradictions in the reported origin of the antiproliferative action of kaempferol, a dietary abundant flavonoid. At >= 10 mu M, kaempferol exposure is, predominantly characterised by cell cycle alterations, notably a significant increase in S-phase and a progressive accumulation in G2-M with 10 and >= 20 mu M kaempferol, respectively. However, a limited but consistent membrane damage is observed across the 1-100 mu M exposure and at 1 mu M occurs devoid from indices of apoptosis which are only consistently observed with >= 10 mu M kaempferol treatment. At the most cytotoxic exposures, multiparametric flow cytometric analysis revealed distinct sub populations of cells. Cells with decreased size, typical of apoptosis and necrosis, possessed heightened caspase-3 activity, decreased anti-apoptotic Bcl-2 expression and changes to membrane asymmetry and integrity. The remaining population had elevated active caspase-3 but no change or a moderate increase in Bcl-2 expression and no plasma membrane alterations. Differentiation was not a significant factor in HL-60 growth inhibition. In conclusion, kaempferol-induced growth inhibition is dominated by cell cycle changes but involves a limited cytotoxicity, which we propose results from a membrane damage centred as well as an apoptotic process. This heterogeneity of response may confound the disease-preventative role and pharmacological application of this flavonoid.