THE IMPLICATION OF PLC-?2 IN GASTRIC CANCER MGC80-3 CELLS TREATED BY TPA / 解剖学报

ABSTRACT

Objective To study the implication of phosphoinositide-specific phospholipase C?2(PLC-?2) in gastric cancer MGC80-3 cells treated by TPA.Methods The effect of 12-O-tetradecanoylphorbol-1,3-acetate(TPA) on gastric cancer MGC80-3 cells was detected and analyzed by DAPI staining under fluorescence microscope.The effect of TPA on the expression level of PLC-?2 protein was detected by Western blotting,when nuclear and cytoplasmic protein fractions were prepared through lysis of cell and centrifugation.Localization and translocation of PLC-?2 were observed under laser scanning confocal microscope with immune-fluorescence technique.After MGC80-3 cells were pretreated by U73122(PLC-?2 inhibitor),the effect of TPA on PLC-?2 was detected by Western blotting and laser scanning confocal microscope with immunefluorescence technique,and the effect of U73122 on MGC80-3 cells followed by TPA was observed and analyzed by DAPI staining under fluorescence microscope. Results TPA induced the apoptosis of gastric cancer MGC80-3 cells.Meanwhile,TPA increased the expression level of PLC-?2 protein and induced its translocation from cytoplasm to nucleus.U73122 inhibited the effect of TPA on the expression level of PLC-?2 protein,but it did not affect on the cell apoptosis induced by TPA.However,the translocation of PLC-?2 protein induced by TPA was not inhibited by its inhibitor(U73122).Conclusion Although TPA could increase the expression level of PLC-?2 protein,it is not directly related to the cell apoptosis induced by TPA.However,its translocation might be related with the apoptosis of gastric cancer.