Effect Of The Effective Extracts And Active Components From Vitex Negundo Seed On The Growth Of MCF-7 Cell Line

Objective: To Screen the effective extracts from 6 species of the extracts from Vitex negundo seed for inhibiting the growth in human breast cancer and to investigate the effect of EVn-50 on inhibiting the growth in human breast cancer in vitro and in vivo, and to determinate the effect of 4 species of lignans from the acetoacetate extract of Vitex negundo seed (comp6, comp7, comp8, comp10) on the viability of human breast cancer MCF-7 cells.Methods: Human breast cancer MCF-7 cells were cultured in vitro. The viability of human breast cancer MCF-7 cells was determinated by MTT assay. Nucleic acid synthesis of MCF-7 cells was examinated by BrdU incorporate assay. The growth of MCF-7 cells was detected using the plate colony formation method. Trial of human breast cancer xenografts in nude mouse medol was used to evaluate the efficacy of Evn-50 on human breast cancer. The volume and weight of xenografts in nude mice were measured. HE staining was utilized to observe the pathological lesion of the xenografts .Result: 6 species of the extracts of Vitex negundo seed (EVn-10,EVn-20, EVn-30 ,EVn-40 ,EVn-50 ,EVn-60)significantly inhibited viability of MCF-7 cells, in a concentration- dependent manner, and their IC50 are 224.06μg/ml,32.67μg/ml,39.34μg/ml,45.90μg/ml, 0.89μg/ml,36.96μg/ml respectively, and which TAX is 0.81μg/ml.BrdU incorporate assay showed that EVn-50 inhibited nucleic acid synthesis of MCF-7 cells in a concentration- dependent manner, and their inhibition ratio is 12%,25 %,34 %,55 %,70% respectively. The plate colony formation method showed that EVn-50 inhibited the growth of MCF-7 cells in a concentration- dependent manner, and their inhibition ratio is 24%,39 %,57 %,65 %,79 %respectively. Trial of human breast cancer xenografts in nude mouse medol showed that Evn-50 significantly inhibited the growth of human breast cancer xenografts in nude mice, in a dose-dependent manner. The weight of xenografts in nude mouse treated with EVn-50 5mg/kg, 10mg/kg and 20mg/kg significantly reduced compared with control group, and their inhibition ratio is 54%,55% and 63%(P <0.001) respectively. However, TAX (10mg/kg) is 66% (P <0.001), TAM (10mg/kg) is 57% (P <0.01). HE staining showed that the cellular necrosis and apoptosis in the xenograft of human breast cancer cells in nude mouse after 16th day of treatment with EVn-50. 4 species of lignans from the acetoacetate extract of Vitex negundo seed(0.1,0.3,1.0,3.0,10.0μg/ml) significantly inhibited viability of MCF-7 cells in a concentration-dependent manner, and their IC50 are 13.16μg/ml(Comp 6) ,37.03μg/ml(Comp 7),3.03μg/m(Comp 8),19.87μg/ml(Comp 10),and which DDP is 10.73μg/ml,EVn-50 is 3.14μg/ml . Comp 8 had more effective than EVn-50.Conclusion:1. EVn-50 inhibits the growth of MCF-7 cell line in vitro and in vivo, and it is the effective extract of Vitex negundo seed for human breast cancer therapy .2. Lignans from the acetoacetate extract of Vitex negundo seed compound 8 had more effective on inhibiting the viability of human breast cancer MCF-7 cells, and it may be one active component of EVn-50.