Pyrazolone copper complex induces apoptosis in BEL-7404 hepatocellular carcinoma cells through extrinsic and intrinsic signaling pathways / 中国药理学通报

ABSTRACT

Aim To investigate the mechanism of apoptosis induced by novel pyrazolone copper complex (P-FAH-Cu-bpy) on human hepatoma cell line BEL-7404. Methods P-FAH-Cu-bpy was prepared by solution method. The crystal structure of P-FAH-Cu-bpy was determined by X-ray single crystal diffraction. The effect of P-FAH-Cu-bpy on the viability of BEL-7404 cells in vitro was detected by MTT assay. The morphology and apoptotic karyotype changes of BEL-7404 cells were observed by inverted microscope and Hoechst 33258 staining, respectively. The effects of P-FAH-Cu-bpy on apoptosis, cell cycle, R O S, mitochondrial membrane potential were detected by flow cytometry. Cell migration was detected by scratch test. The effects of P-FAH-Cu-bpy on apoptosis, migration and invasion-related protein expression in BEL-7404 cells were assessed by Western blot. Results P-FAH-Cu-bpy as a pentacoordinate copper (II) complex with a twisted tetragonal pyramidal configuration suppressed cell proliferation in a dose- and time-dependent manner, induced apoptosis, arrested the cell cycle in the G2/M phase, decreased the mitochondrial membrane potential, and increased the levels of intracellular ROS. The expression levels of Bax, cytochrome C, Cleavedcaspase-3/9/8 and Cleaved-PARP were up-regulated, while the expression levels of caspase-7 and Bcl-2 were down-regulated; cell migration and invasion were inhibited. Conclusion P-FAH-Cu-bpy suppresses BEL-7404 cell growth through both extrinsic and intrinsic apoptosis pathways.