Studies Of Extracts From Inonotus Obliquus On Anti-proliferation And Inducing Apoptosis On BGC-823 Human Gastric Cancer Cell Line

OBJECTIVE To observe the effects of anti-proliferation and inducing apoptosis on BGC-823 human gastric cancer cell line induced by extracts from Inonotus obliquus and the protein expressions of apoptosis-related gene in vitro, and provide the experimental evidence for Inonotus obliquus extracts as a new anti-tumor fungus drug.METHODS BGC-823 human gastric cancer cell line was cultured in vitro. MTT assay was used to determine the effect of extracts from Inonotus obliquus extracted by different methods on BGC-823 human gastric cancer cells growth inhibitory. Morphological changes of the apoptosis of BGC-823 gastric cells were observed by using inverted, light and fluorescent microscope, and the ultrastructure of BGC-823 cells was observed by using transmission electronic microscope. Apoptotic rate was calculated with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and apoptotic half-quantitative method through AO/EB double fluorescent staining. Flow cytometry was used to detect cell cycle distribution of BGC-823 cells and apoptotic rate. Furthermore, the protein expressions of Ki-67 and the apoptosis-related gene such as bax, bcl-2, caspase-3 and caspase-9 were detected by using immunocytochemistry before and after exposure to aqueous extract from Inonotus obliquus.RESULTS 1) The MTT test showed that BGC-823 gastric cells exposure to aqueous extract from Inonotus obliquus, petroleum ether extract from Inonotus obliquus and acetic ether extract from Inonotus obliquus at final concentrations of 10~80μg/ml were inhibited for 12h, 24h, 48h and 72h in dose and time dependent manner. The MTT test showed the inhibitory rate was 51%, 60% and 62%, respectively at 80μg/ml final concentration for 48h. The effect of petroleum ether and acetic ether extracts from Inonotus obliquus inhibiting growth of human BGC-823 gastric carcinoma cell line in vitro was more marked than that of aqueous extract from Inonotus obliquus. 2) The BGC-823 human gastric cancer cells in experimental group by HE staining...