Role Of Fas Signaling Pathway In DADS-induced Apoptosis Of MSB-1 Cells

Objective: To explore the mechanism of garlic extract diallyl disulfide(DADS)-induced apoptosis of lymphoproliferative tumor cell line MDCC-MSB-1 caused by chicken Marek’s disease virus(MDV),from exogenous Fas /Fas L Apoptosis signaling pathway explores the mechanism of DADS-induced apoptosis in MSB-1 cells,which can provide a scientific basis for the treatment of tumors caused by Marek’s disease virus,as well as the development of new drugs and the treatment of human tumor diseases caused by DNA viruses guess.Methods: In this study,MDCC-MSB-1 cells were used as the test object,and the CCK-8 method was used to initially explore the effect of equal gradient concentration of DADS on the cell viability of MSB-1 cells after 24 h,48 h and 72 h respectively,and confirm the test low,medium and high doses Group concentration.The effect of DADS on the morphology of MSB-1 cells: observation by Hoechst 33342 fluorescence staining.DADS-induced apoptosis in MSB-1 cell apoptosis mechanism of Fas/Fas L signaling pathway m RNA: fluorescence quantitative q PCR detection.DADS induces MSB-1 cells to undergo apoptosis of Fas / Fas L signaling pathway,a key enzyme activity: Caspase-8 activity detection kit.Apoptosis status of MSB-1 cells after DADS treatment for 24 h: Annexin V-FITC / PI double staining flow cytometry.Results: 40-240 μmol / L DADS inhibited the viability of MSB-1 cells in a time-and dose-dependent manner,and determined the concentration of low,medium and high dose groups(80 μmol / L,160 μmol / L,240 μmol / L).After 24 hours of DADS treatment on MSB-1 cells,the cell morphology changed from the round and transparent cell nucleus to the disappearance of the nuclear membrane,and the chromosomal DNA was degraded into multiple fragments.The expression of Fas,FADD,Caspase-8,Bid and Caspase-3 m RNAs of Fas / Fas L signaling pathway-associated Fas,FADD,Caspase-8,Bid,and Caspase-3 after MSDS-1cells were treated with DADS at concentrations of 80 μmol / L,160 μmol / L,and 240 μmol/ L.Drug concentration-dependent trend.After 24 h of DADS at 80 μmol / L,160 μmol / L,and 240 μmol / L on MSB-1 cells,the percentage increase of Caspase-8 protease activity showed a concentration-dependent trend.After 24 h of MSB-1 cells treated with DADS at concentrations of 80 μmol / L,160 μmol / L,and 240 μmol / L,the number of viable cells decreased,and most of them were in the early stage of apoptosis.Conclusion: Diallyl disulfide can inhibit the proliferation of chicken Marek’s disease lymphoma MSB-1 cells,and can induce the apoptosis of MSB-1 cells.Among them,the diallyl disulfide with a concentration of 80-240μmol / L induced the apoptosis of MSB-1cells in a time-and dose-dependent manner.The mechanism of inducing apoptosis is the coexistence of mitochondrial endogenous apoptosis pathway and Fas / Fas L exogenous apoptosis pathway.