Induction of apoptosis by osthole in HL-60 cells and the molecular mechanism research / 国际肿瘤学杂志

ABSTRACT

Objective To detect the effect of osthole on proliferation and apoptosis of HL-60 cells and its molecular mechanism.Methods HL-60 cells proliferation was measured through the CCK8 assay method.The cell morphological changes were observed by Hoechst33342 staining after 8 h of drug effect.Induction of apoptosis was determined by flow cytometry and fluorescent microscopy.Expressions of Bcl-2 and Bax mRNA were evaluated by RT-PCR,and the expressions of cleaved caspase-3,caspase-8,caspase-9,Fas and FasL were evaluated by using western bolt assay.Results Osthole could inhibit the proliferation of HL-60 cells,the maximum inhibiting rate was (90.7 ±4.5)％,F =138.46,P =0.000; the apoptosis rate was 33.6％,F =27.75,P =0.006.The changes of apoptosis of cells and nucleus were shown in cell morphological observation.Osthole affected the decrease of the mRNA levels of Bcl-2 and the increase of the Bax mRNA levels via a dosedependent manner(F =210.12,P =0.000).Western blotting demonstrated that osthole could lead to the increase of the expression levels of cleaved caspase-3,caspase-8,caspase-9,Fas and FasL in the HL-60 cell line via a time-dependent manner.Conclusion Data suggests that osthole inhibits proliferation and induces apoptosis of HL-60 cells through mitochondria-dependent pathway and death-receptor pathway.