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AIM:To investigate the roles of PLCγ2 and PKCα in TPA-induced apoptosis of gastric cancer cells.METHODS:Human gastric cancer cell line MGC80-3 wasused.Protein expression levels of PLCγ2 and PKCα weredetected by Western blot.Protein localization of PLCγ2 andPKCα was shown by immunofluoscence analysis under laser-scanning confocal microscope.Apoptotic morphology wasobserved by DAPI fluorescence staining,and apoptotic indexwas counted among 1 000 cells randomly.RESULTS:Treatment of gastric cancer cells MGC80-3 withTPA not only up-regulated expression of PLC-γ2 protein,but also induced PLC-γ2 translocation from the cytoplasm tothe nucleus.However,this process was not directly associatedwith apoptosis induction.Further investigation showed thatPKCa translocation from the cytoplasm to the nucleus wascorrelated with initiation of apoptosis.To explore the inevitablelinkage between PLC-γ2 and PKCα during apoptosis induction,PLC inhibitor U73122 was used to block PLC-γ2 translocation,in which neither stimulating PKCα translocation nor inducingapoptosis occurred in MGC80-3 cells.However,when U73122-treated cells were exposed to TPA,not only PLC-γ2,but alsoPKCα was redistributed.On the other hand,when cells weretreated with PKC inhibitor alone,PLC-γ2 protein was stilllocated in the cytoplasm.However,redistribution of PLC-γ2protein occurred in the presence of TPA,no matter whetherPKC inhibitor existed or not.CONCLUSION:PLC-γ2 translocation is critical in transmittingTPA signal to its downstream molecule PKCα.As an effector,PKCα directly promotes apoptosis of MGC80-3 cells.Therefore,protein translocation of PLCγ2 and PKCα is criticalevent in the process of apoptosis induction.
AIM: To investigate the roles of PLCγ2 and PKCα in TPA-induced apoptosis of gastric cancer cells. METHODS: Human gastric cancer cell line MGC80-3 wasused. Protein expression levels of PLCγ2 and PKCα weredetected by Western blot. Protein localization of PLCγ2 and PKCα was Observation by immunofluoscence analysis under laser-scanning confocal microscope. Apoptotic morphology wasobserved by DAPI fluorescence staining, and apoptotic index was counted among 1 000 cells randomly .RESULTS: Treatment of gastric cancer cells MGC80-3 withTPA not only up-regulated expression of PLC- γ2 protein, but also induced PLC-γ2 translocation from the cytoplasm tothe nucleus. Although this process was not directly associated with apoptosis induction. Further investigation showed that PKCa translocation from the cytoplasm to the nucleus wascorrelated with initiation of apoptosis. To explore the inevitable linkage between PLC- γ2 and PKCα during apoptosis induction, PLC inhibitor U73122 was used to block PLC-γ2 translocation, in which neither stimulating PKCα translocation nor inducingapoptosis occurred in MGC80-3 cells. When, when U73122-treated cells were exposed to TPA, not only PLC-γ2, but also PKCα was redistributed. On the other hand, when cells weretreated with PKC inhibitor alone , PLC-γ2 protein was stilllocated in the cytoplasm. However, redistribution of PLC-γ2protein occurred in the presence of TPA, no matter whether PKC inhibitor existed or not. CONCLUSION: PLC-γ2 translocation is critical in transmitting TPA signal to its downstream molecule PKCα. As an effector, PKCα directly promotes apoptosis of MGC80-3 cells. Beforefore, protein translocation of PLCγ2 and PKCα is criticalent in the process of apoptosis induction.