Diallyl Disulfide Mediated Autophagy In Leukemia K562Cell Through Up-regulation Of Beclin1

Objective: To investigate the possibility of Beclin1mediated autophagy inK562cell induced by diallyl disulfide (DADS).Methods: K562cells were cultured in K562cell culture systems;K562cellswere treated with DADS(10mg/L,20mg/L,40mg/L,80mg/L) for12hours and40mg/LDADS treatment with K562cells for indicated time(3hours,6hours,12hours,24hours) the autophagic vacuoles(AV) were observed byfluorescence microscope through monodansylcadaverin(MDC) staining; K562cellswere treated with40mg/L DADS for12hours,they ultrastructure were observed bytransmission election microscopy(TEM);and the rates of autophagy were analyzed byflow cytometry after treated with different concentrations of DADS;The expressionlevel of Beclin1mRNA and Protein were measured by RT-PCR and Westernblotting after treated with40mg/L DADS for12hours.Results:Compared with the blank reference group and the vehicle referencegroup, the autophagic vacuoles determined by MDC staining fluorescence microscopewere increased significantly in different concentration of DADS groups and indifferent duration groups, the most autophagic vacuoles were observed in40mg/LDADS group and40mg/LDADS treatment for12h group;After treatmentwith40mg/L DADS for12hours, large amount of autophagosomes were observed incytoplasm of K562cell by TEM;The autophagy rate of the blank reference group,the vehicle reference group， were(0.97±0.19)%、(1.05±0.20)%（P>0.05）)respectively,and10mg/L、20mg/L、40mg/L、80mg/LDADS group, were(10.11±0.08)%、(21.55±0.34)%、(34.36±2.15)%、(19.20±0.95)%respectively,compared with control group(P <0.05). The expression level of Beclin1mRNA andProtein were significantly increased after treated with40mg/L DADS for12hours，compared with control group(P <0.05),while the Blank reference group compared with the Vehicle reference group（P>0.05）.Conclusion:DADS can induce autophagy in leukemia K562cell,it may berelated to the up-regulation of Beclin1expression.