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AIM:To investigate the roles of Fas signaling pathway invitamin E succinate-induced apoptosis in human gastriccancer SGC-7901 cells.METHODS:Human gastric cancer SGC-7901 cells weretreated with VES at 5,10,20 mg·L~(-1),succinic acid and vitaminE as vehicle control and condition media only as untreated(UT)control.Apoptotic morphology was observed by DAPIstaining.Western blot analysis was applied to measure theexpression of Fas,FADD and caspase-8 proteins.After thecells were transiently transfected with Fas and FADDantisense oligonucleotides,respectively,caspase-8 activitywas determined by flurometric method.RESULTS:The morphologically apoptotic changes wereobserved alter VES treatment by DAPI staining.23.7 % and89.6 % apoptosis occurred alter 24 h and 48 h of 20 mg·L~(-1)VES treatment,respectively.The protein levels of Fas,FADDand caspase-8 were evidently increased in a dose-dependentmanner alter 24 h of VES treatment.The blockage of Fas bytransfection with Fas antisense oligonucleotides obviouslyinhibited the expression of FADD protein.After SGC-7901cells were transfected with Fas and FADD antisenseoligonucleotides,caspase-8 activity was obviously decreased(P<0.01),whereas Fas blocked more than FADD.CONCLUSION:VES-induced apoptosis in human gastriccancer SGC-7901 cells involves Fas signaling pathwayincluding the interaction of Fas,FADD and caspase-8.
AIM: To investigate the roles of Fas signaling pathway invitamin E succinate-induced apoptosis in human gastriccancer SGC-7901 cells.METHODS:Human gastric cancer SGC-7901 cells weretreated with VES at 5,10,20 mg·L -1 ,succinic acid and vitaminE as vehicle control and condition media only as untreated(UT)control.Apoptotic morphology was observed by DAPIstaining.Western blot analysis was applied to measure the expression of Fas,FADD and caspase-8 proteins.After the cells were transiently transfected with Fas and FADDantisense oligonucleotides, relevantly, caspase-8 activitywas determined by flurometric method.RESULTS: The morphologically apoptotic changes wereobserved alter VES treatment by DAPI staining. 23.7% and 89.6 % apoptosis occurred alter 24 h and 48 h of 20 mg·L~ (-1) VES treatment, variously.The protein levels of Fas,FADDand caspase-8 were evidently increased in a dose-dependentmanner alter 24 h of VES treatment.The blockage of Fas bytransfection with Fas antisense oligonucleotid Es significantlyinhibited the expression of FADD protein.After SGC-7901cells were transfected with Fas and FADD antisenseoligonucleotides, caspase-8 activity was clearly decreased(P<0.01), whereas Fas blocked more than FADD.CONCLUSION:VES-induced apoptosis in human gastriccancer SGC -7901 cells Fas signaling pathway including the interaction of Fas, FADD and caspase-8.