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AIM:To investigate the roles of c-Jun N-terminal kinase (JNK)signaling pathway in vitamin E succinate-induced apoptosisin human gastric cancer SGC-7901 cells.METHODS:Human gastric cancer cell lines (SGC-7901)were treated with vitamin E succinate (VES) at 5,10,20 mg/L.Succinic acid and vitamin E were used as vehicle controlsand condition medium only as an untreated (UT) control.Apoptosis was observed by 4′,6-diamidine-2′-phenylindoledihydrochloride (DAPI) staining for morphological changesand by DNA fragmentation for biochemical alterations.Western blot analysis was applied to measure the expressionof JNK and phosphorylated JNK.After the cells were transientlytransfected with dominant negative mutant of JNK (DN-JNK) followed by treatment of VES,the expression of JNKand c-Jun protein was determined.RESULTS:The apoptotic changes were observed after VEStreatment by DNA fragmentation.DNA ladder in the 20 mg/LVES group was more clearly seen than that in 10 mg/L VESgroup and was not detected following treatment of UTcontrol,succinate and vitamin E.VES at 5,10 and 20 mg/Lincreased the expression of p-JNK by 2.5-,2.8- and 4.2-fold,respectively.VES induced the phosphorylation of JNKbeginning at 1.5 h and produced a sustained increase for24 h with the peak level at 12 h.Transient transfection ofDN-JNK blocked VES-triggered apoptosis by 52%.DN-JNKsignificantly increased the level of JNK,while decreasingthe expression of VES-induced c-Jun protein.CONCLUSION:VES-induced apoptosis in human gastriccancer SGC-7901 cells involves JNK signaling pathway viac-Jun and its downstream transcription factor.
AIM: To investigate the roles of c-Jun N-terminal kinase (JNK) signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells. METHODS: Human gastric cancer cell lines Eucate (VES) at 5, 10, 20 mg / L.Succinic acid and vitamin E were used as vehicle controls and condition medium only as an untreated (UT) control. Apoptosis was observed by 4 ’, 6-diamidine- phenylindoledihydrochloride (DAPI) staining for morphological changes and by DNA fragmentation for biochemical alterations. Western blot analysis was applied to measure the expression of JNK and phosphorylated JNK. After the cells were transiently transfected with dominant negative mutant of JNK (DN-JNK) followed by treatment of VES , the expression of JNK and c-Jun protein was determined .RESULTS: The apoptotic changes were observed after VES treatment by DNA fragmentation. DNA ladder in the 20 mg / LVES group was more clearly seen than that in 10 mg / L VESgroup and was not detec ted following treatment of UTcontrol, succinate and vitamin E. VES at 5, 10 and 20 mg / Lincreased the expression of p-JNK by 2.5-, 2.8- and 4.2-fold, respectively. VES induced the phosphorylation of JNKbeginning at 1.5 h and produced a sustained increase for 24 h with the peak level at 12 h. Transient transfection of DN-JNK blocked VES-triggered apoptosis by 52% .DN-JNKsignificantly increased the level of JNK, while decreasingthe expression of VES-induced c-Jun protein.CONCLUSION : VES-induced apoptosis in human gastriccancer SGC-7901 cells involves JNK signaling pathway viac-Jun and its downstream transcription factor.