Relationship between PTEN-Akt pathway and 8-nitrochrysin induced apoptosis and inhibition of the proliferation of HL-60 cells

OBJECTIVE To observe the inhibition effect of 8-nitrochrysin on the proliferation of HL-60 cell and induction of HL-60 cell apoptosis,and investigate the action mechnism.METHODS HL-60 cell line was cultured in vitro and the proliferative activity of HL-60 cells treated with 5,7-dihydrox-8-nitrochrysin was measured using 3-(4,5-dimethylthiazol –2-yl)-2,5-diphenytertrazolium blue(MTT) colorimetric assay.The apoptosis rate of HL-60 cell line induced by NOChR was analyzed using flow cytometry(FCM) with PI staining.DNA ladder bands were observed by DNA agarose gel electrophoresis.The influence of NOChR on the PTEN and p-AKt protein expression of HL-60 cells was analyzed with western blot.RESULTS The proliferation of HL-60 cells was markedly inhibital by NOChR in a concentration-dependent manner,and the IC50 was 1.34 μmol.Flow cytometry after PI staining indicated that apoptosis in HL-60 cells was induced by NOChR in a time and concentration dependent manner.Western blot indicated that NOChR can efficiently up-regulated PTEN and down-regulated p-AKt protein expression of HL-60 cells in a concentration and time dependent manner,and the pre-incubation of GW9662,blocker of PPARγ,can antagonize the up-regulating effect of 8-nitrochrysin on PTEN protein expression of HL-60 cells.CONCLUSION NOChR induced apoptosis of HL-60 cells may be associated with activation of PPARγ,up-regulation of PTEN and inhibition of p-AKt by 8-nitrochyrsin.