Study On The Anti-tumor Activity Of Abrus Cantonensis Extraction

Objective: This study was to investigate the inhibitory effect of active components extract from Abrus cantonensis with four different polar solvents on SGC-7901 gastric cancer cell,BEL-7404 hepatoma cells,MCF-7 breast cancer cells in vitro,and than screened out Abrus cantonensis extract has the anti-tumor function in vitro.Safety evaluation of Abrus cantonensis extract was evaluated in vivo.To establish subcutaneous transplanted model group of SGC-7901 gastric cancer cell,BEL-7404 hepatoma cells,MCF-7 breast cancer cells in nude mice and to explore the extracts of Abrus cantoniensis with antitumor effect in vivo.And finally,sift out the Abrus cantonensis extract has the anti-tumor function in vivo and in vitro,then it is a good basis and direction for further separation of monomer compounds have antineoplastic activity from Abrus cantonensis extracts.Methods:(1)The SGC-7901,BEL-7404,MCF-7 cancer cells were used as cell mode,inhibitory effect of Abrus cantoniensis extract on cell proliferation was measured by the method of CCK-8,observing the morphology of the cells under inverted phase contrast microscope.(2)The number of apoptotic cell was calculated by hoechst33258 fluorescent staining,the condition of the cell cycle and apoptosis were analyzed by flow cytometry.(3)Acute toxicity experiment was used to evaluate the safety of Abrus cantonensis extractions.(4)Tumor model group of SGC-7901,BEL-7404,MCF-7 mice were evaluated the effect of Abrus cantonensis extract on inhibition rate of tumor,tissue necrosis was observed by hematoxylin-eosin(HE)staining.Proteins expressionof Bax and Bcl-2 in BEL-7404 tumor tissue were measured by immunohistochemistry.Results:(1)The result of CCK-8 showed the petroleum ether,ethyl acetate,butyl alcohol,water extract from Abrus cantoniensis inhibited the proliferation of SGC-7901,BEL-7404,MCF-7 cancer cells in a dose-dependent manner,except the petroleum ether extract,the others at least had 50% mortality when the drug concentration was about 0.7 mg/m L and the ethyl acetate extract was most active.(2)Hoechst 33258 fluorochrome staining and Annexin V-FITC/PI double staining showed that the ethyl acetate extract could pro-mote BEL-7404 cells apoptosis(P<0.001),the apoptosis rate was gradually increased with the drug concentration increase.Propidium iodide(PI)staining method indicated that there was more cells in G0/G1 phase and less cells in G2/M phase after treated with different concentrations of the ethyl acetate extract,when compared with the blank control group and the above differences were statistically significant(P<0.05).(3)There was no significant difference of the weight change of mice between the control group with the experiment group.There is not toxic symptoms and deaths mices.The shape,colour,size of major organs in mice were not found obvious abnormal changes after dissected.Median lethal dose of ethyl acetate,n-butanol and water extract of Abrus cantoniensis in mice were more than 16080,27600,35360 mg/kg respectively.It can be judged that ethyl acetate,n-butanol and water extract were non-toxic,according to the value of acute toxicity dose classification standard table.The results showed that ethyl acetate,n-butanol and water extract of Abrus cantoniensis have no toxic effect on mice.(4)The studies indicated that the ethyl acetate,butyl alcohol,water extract from Abrus cantoniensis have anti-tumor effect in vivo and compared with model group there was a significant difference statistically(P<0.05),along with extract density increasing,the ability of anti-tumor activity also is strengthened.And the ethyl acetate was most active,after treatment with the ethyl acetate extract,the growth inhibitory rates of SGC-7901,BEL-7404,MCF-7 cancer cells xenograft in nude mice were 43.64%,47.35%?and 29.53%.The results of HE staining showed that: each extract dosage groups showed various degrees of SGC-7901,BEL-7404,MCF-7 necrosis,nuclear enrichment,fragmentation and vanishment,the group of the ethyl acetate extract were more serious.Immunohistochemical results of the integral optical density of Bax in high-dose,midle-dose ethyl extracts of Abrus cantonensis group was higher than that in the model group,while the integral optical density of Bcl-2 was lower than that in the model group,the difference was statistically significant(P<0.05);and there was no significant difference in the integrated optical density(IOD)of Bax,Bcl-2 between the low-dose ethyl extracts of Abrus cantonensis group and the model group(P>0.05).Conclusion:(1)The activity sequence of Abrus cantonensis extraction inhibiting cell proliferation of the tumour cell in vitro was that: the petroleum ether extract> the ethyl acetate extract>the butyl alcohol extract>the water extract;(2)The ethyl acetate extract of Abrus cantonensis can induce BEL-7404 cells apoptosis and blocke the cell cycle in G0/G1 phase;(3)The results of acute toxicity tests show that ethyl acetate,n-butanol and water extract had no toxic effect on mice;(4)The petroleum ether,ethyl acetate,butyl alcohol,water extract from Abrus cantoniensis could inhibits the growth of SGC-7901,BEL-7404,MCF-7 cancer cells transplantation tumor in nude mice,the ethyl acetate extract of Abrus cantonensis has the funtions of promonting Bcl-2 and inhibiting Bax proteinic expression.