DADS Through PI3K/Akt Signaling Pathways Induced Leukemia K562 Cells Apoptosis

Objective:The aim of this study was to explore the possible mechanism of K562 cell treated with DADS in the PI3K/Akt signaling pathway,and to provide experimental basis for further clinical application. Methods:K562 cells were treated with DADS at different concentrations of 10 to 80mg/L for 48 h when they were in the logarithmic growth phase, The morphologic changes of leukemia K562 were observed by Inverted microscope?AO/EB stain. Flow cytometry(FCM) detect the apoptosis rate of K562 cells treated with DADS for 48 h. The expression of Akt, P-Akt and Caspases-3 protein was detected by Western blotting, setting up blank and solvent control group as a reference. Results:1. Inverted microscope demonstrated that treatment of K562 cells with DADS for 48 h showed the characteristic morphological changes of apoptosis, such as: Smaller volume, Condensed chromatin, Cell membrane outstanding.2.AO/EB stain demonstrated that the gradually increase of the concentration of DADS, Nuclei in a solid-like or bead-like condensation and chromatin condensation stained orange of apoptosis cells increased, concentration of 40mg/L DADS group was the most significant.3.Annexin V-FITC/PI double staining showed that apoptosis rate increased gradually with increasing concentration of DADS, the apoptosis rates were(9.4 ± 0.87)%,(15.1 ± 0.40)%,(27.4 ± 2.60)%,(18.1 ± 1.20)%, compared with the control group of(3.2± 0.92)% and(2.9±0.26)% had significant difference(P<0.05).4. With the increase of the concentration of DADS, the expression of Akt protein in treated group with DADS were have no significant difference compared with the control group(P>0.05); Akt protein phosphorylation levels were gradually decreased in 10, 20, 40mg/L DADS group, and sho- wed a certain dose dependence, the difference compared with the control group was statistically significant(P<0.05), when the DADS concentration was 40mg/L, its relative expression was the lowest, and the difference was most significant(P<0.05); The expression of caspases-3 was just contrary to the expression of P-Akt protein,the most obvious of caspases-3,s expression at 40mg/L(P<0.05). ConclusionThe related molecular mechanism is that DADS induce apoptosis of K562 cells are through down-regulate PI3K/Akt signaling pathway expression and up-regulate Caspases-3 expression.