Effects And Mechanisms Of Co-ASS On The Proliferation And Apoptosis Of KB Cells

Objective: To investigate the effects of Co-ASS on the proliferation and apoptosis of human oral squamous KB cell, and explore the underlying mechanisms.Materials and Methods: KB cells were treated with different concentrations of Co-ASS in vitro. Cell viability was determined by methyl thiazolyl tetrazolium(MTT) and colony formation assay. Cell cycle were analyzed by flow cytometry. Apoptosis was measured by Hoechst 33342 fluorescence staining and flow cytometry. Intracellular reactive oxygen species(ROS) was tested with DCFH-DA probe. Mitochondrial membrane potential(MMP) was quantified by staining with JC-1 probe. Expression of P53、Bcl-2、Bax、Cox-2 were detected by western blot.Result:(1) The colony formation efficiency and the ability of cell proliferation of KB cells were both reduced after being exposed to Co-ASS in a dose-dependent manner(P<0.05).(2) Flow cytometry analysis showed that the cell cycle of KB was arresred in G2/M phase and the apoptosis rate was increased. After Hoechst 33342 staining, the cells showed apoptotic morphological changes such as the cell nucleus changes like pyknosis and karyorrhexis and dense blue fluorescent particles can be observed.Compared with control group,the difference was statistically significant(P<0.05).(3) The ROS levels of KB cells was increased and the MMP was decreased after treatment with Co-ASS. Western blotting analysis showed that the expression of P53、Bax were significantly upregulated and Bcl-2、COX-2 were downregulated. Compared with control group,the difference was statistically significant(P<0.05).Conclusion: Co-ASS could inhibit the proliferation and induce the apoptosis of KB cells in a dose-dependent manner, which may be correlated with the increase of ROS and down regulation of COX-2.