Mechanism Of G2/M Arrest Induced By DADS In MDCC-MSB-1 Cells

Objective:To investigate the mechanism of garlic extract diallyl disulfide(DADS)inducing proliferation inhibition of lymphoproliferative tumor cell line MDCC-MSB-1 induced by Marek's virus(MDV).To study the blocking mechanism of DADS on MDCC-MSB-1 cells in the direction of cellular pathway can provide scientific basis for preventing the occurrence of Marek's disease and for speeding up the development of new drugs.Methods:In this study,MDCC-MSB-1 cells were treated with DADS(50 ?mol/L,100 ?mol/L,150 ?mol/L)for 24 hours,and the control group and the drug group were treated with different concentration gradient drug DADS(50 ?mol/L,100 ?mol/L,150 ?mol/L)for 24 hours.To explore the mechanism of G2/M phase arrest induced by DADS in MDCC-MSB-1 cells.(1)Effects of DADS on the morphology of MDCC-MSB-1 cells: observed under OLYMPUS fluorescence microscope.(2)Effect of DADS on G2/M phase arrest of MDCC-MSB-1 cells: cell cycle detection kit assay.(3)Effects of DADS on the related proteins in G2/M arrest mechanism of MDCC-MSB-1 cells: Western Blot was used to detect the expression of CDK1,CyclinB1,WEE1,MYT1,P53 and P21 proteins.Results:(1)After treated with DADS for 24 hours,the morphology of the cells changed obviously from round clear nucleus to blurred cell membrane boundary,and the higher the drug concentration,the more obvious the change of cellular organelle nucleus fragmentation.(2)The proliferation of MDCC-MSB-1 cells was significantly inhibited in G2/M phase after 24 hours of DADS treatment with 50 ?mol/L,100 ?mol/L and 150 ?mol/L concentration respectively.With the increase of drug concentration,the inhibition was more obvious and showed a concentration-dependent trend.(3)The expression of CDK1 protein,CyclinB1 protein,P53 protein,WEE1 and MYT1 protein in DADS-treated cells were up-regulated.The expression of P21 protein in three drug concentrations was down-regulated compared with non-drug groups,but there was little difference among the three drug groups.Conclusion:DADS had a certain effect on the morphology of MDCC-MSB-1 cells.Diallyl disulfide(DADS)can induce G2/M phase proliferation arrest of lymphoplastic tumor cell line MDCC-MSB-1 cells induced by MDV.The blocking mechanism is to block cell proliferation in G2/M phase by regulating the expression of related proteins in WEE1/MYT1/CDK1/CyclinB1 pathway and P53/CDK1/CyclinB1 pathway.