Effects of Bruceine D on the proliferation, apoptosis and autophagy of osteosarcoma cells

ZHENG Ying; WANG Gang-yang; CHEN Rui-ling; HUA Ying-qi; CAI Zheng-dong

doi:10.3969/j.issn.2095-252X.2017.06.007

Abstract：Objective To investigate the definite effects of Bruceine D on inhibiting the proliferation of osteosarcoma cells and inducing the apoptosis and autophagy in osteosarcoma cells. Methods The anti-proliferative effects of Bruceine D on 143B HOS cells were detected by CCK8 assay. Colony formation assay was used to detect the colony formation of 143B HOS cells treated with Bruceine D. The Bruceine D's effects of cell cycle arresting on 143B HOS cells were analyzed by flow cytometry. The Bruceine D's effects of inducing apoptosis in 143B HOS cells were investigated by flow cytometry and Western blot assay. We measured the autophagy associated proteins expressed in 143B HOS cells treated with Bruceine D by fluorescence microscope. The Bruceine D's effects of inducing autophagy were also investigated by GFP-LC3 punctate assay. Results ( 1 ) The CCK8 assay showed that the cell viability and number of colonies of 143B HOS treated with Bruceine D were decreased. ( 2 ) A higher ratio of G0 to G1 phase cells in 143B HOS was caused by Bruceine D, which was found by using flow cytometry. ( 3 ) The apoptosis proportion of 143B HOS treated with Bruceine D was increased, which was found by using flow cytometry; The western blot assay showed the expression level of Cleaved PARP was increased in 143B HOS. ( 4 ) The western blot assay showed the expression level of LC3B-II was increased in 143B HOS; The 143B cells treated by Bruceine D displayed a punctate pattern of GFP-LC3 expression, which was found by using fluorescence microscope. Conclusions ( 1 ) Bruceine D inhibits the proliferation of osteosarcoma cells. ( 2 ) Bruceine D induces the apoptosis of osteosarcoma cells. ( 3 ) Bruceine D induces the autophagy of osteosarcoma cells.