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Effect of sinomenine on proliferation and apoptosis of oral squamous cell carcinoma cells, and its underlying mechanism of action
Abstract
Purpose: To investigate the effect of sinomenine on the proliferation and apoptosis of oral squamous cell carcinoma cells (OCSSs), and its underlying mechanism of action.
Methods: CCK-8 method was used to determine the inhibitory effects of different concentrations of sinomenine (50, 100, 200, 400 and 800 ug/mL) on proliferation of CAL-27 cells on treatment for 24 and 48 h. Cell migration was assayed using scratch test, while cell cycle was measured by flow cytometry. Hoechst 33258 fluorescence staining was conducted to determine apoptotic morphology. The effect of sinomenine on the expressions of cyclinD1, cyclinD3, cyclin-dependent kinases-2 (CDK2), Bax, B-cell lymphoma/leukmia-2 (Bcl-2), Caspase-3 and Caspase-9 was determined by western-blotting (WB).
Results: Sinomenine significantly inhibited proliferation and migration of CAL-27 cells in a concentration-dependent manner, but not in a time-dependent fashion. It inhibited the transition from G0/G1 phase to S phase in CAL-27 cells. Sinomenine treatment also caused dispersion of CAL-27 cells. Moreover, the nuclei were pyknotic, and the cells translucent, indicating typical features of apoptotic morphology. Expressions of Bax, caspase 3 and caspase 9 protein were significantly upregulated, while expressions of Bcl-2, cyclinD1, cyclinD3 and CDK2 protein were down-regulated by sinomenine.
Conclusion: Sinomenine induces apoptosis of OSCCs, and inhibits their proliferation and migration via a mechanism associated with up-regulation of Bax, Caspase-3 and Caspase-9, and down-regulation of Bcl-2.