Study On Mechanism And Effect Of Apoptosis And Proliferation By (一)-gossypol On Human Synovial Sarcoma Cells

Object:To study the eftectsofmanrunycin on proliferation and apoptosis of human synovial sarcoma SW-982 cells and elucidate the possible mechanisms. Methods:MTT colorimetric assay was applied to detect the antiproliferation of (-)-gossypol in synovial sarcoma SW-982 cells; Morphological changes in the nuclear chromatin of cells undergoing apoptosis. were determined by Wright-Giemsa staining, Hoechst33258 staining and transmission electron microscopy detection; Flow cytometric assay was performed to evaluate the cell cycle and apoptosis by staining with PI as well as Annexin V/PI; The influence of (-)-gossypol to the expression of apoptosis-associated proteinBcl-2 and Bax were detected by Western blot; Caspase-9 and Caspase-3 activation was analyzed by colorimetric assay. Results:(-)-Gossypol could inhibite the proliferation of synovial sarcoma cells SW-982 in dose and time dependent manner. After treated with 4.0μmol/L (-)-gossypol, Wright-Giemsa staining, Hoechst33342/Pl staining and transmission electron microscopy detection showed t the typical apoptotic features. FCM indicated that after the treatment of (-)-gossypol for 24h, the proportion of the G0/G1 phase cells was increasing accompanied with the concentrations, but the proportions of G2/M and S phases cells decreased. Gossypol blocked cells at G0/G1 phases, this phenomenon demonstrated that (-)-gossypol could regulate the cell cycle of SW-982 cells. FCM detected that the cell early apoptotic rates had a positive correlation relationshipwith the concentration of (-)-gossypol. After treatment of (-)-gossypol at the concentration of 4.0μmol/L for 24h, the expression of apoptosis-associated protein Bcl-2 of SW-982 cells had a positive correlation relationshipwith the concentration of (-)-gossypol. the expression of apoptosis-associated protein Bax of SW-982 cells had a negative correlation relationshipwith the concentration of (-)-gossypol. Caspase-9 and Caspase-3 activation was confirmed at the concentration of 4.0μmol/L treated by (-)-gossypol. When SW-982 cells were exposed to (-)-gossypol for 6h, Caspase-9 activation was irritated and the activity reached the peak at 12h, then the activity showed a descending tendency. When SW-982 cells were exposed to (-)-gossypol for 6h, Caspase-3 activation was irritated and the activity reached the peak at 18h, then the activity showed a descending tendency.Conclusion:In this study, we demonstrated that (-)-gossypol was able to inhibit the proliferation and induce apoptosis of synovial sarcomas cells SW-982, the mechanisms might include down regulation of Bcl-2 protein, up regulation of Bax protein and Caspase-9 and Caspase-3 activation. (-)-Gossypol acetate induced SW-982 cells apoptosis by the mitochondrial pathway.