The Effect Of Calycosin On The Proliferation Of Human Breast Cancer Cells And Its Mechanism

Objective: Calycosin is one of the major components of isoflavones,it is a natural plant estrogen,its structure is similar to the structure of estrogen.Calycosin has some certain effects respectively in antiviral and anti cardiovascular disease,especially in the inhibition of human malignant tumor proliferation,which attracted much attention.In our previous studies,we concluded that calycosin inhibited growth in breast cancer cells,but mechanism is not clear.In our studies,we further to study the mechanisms of effects of calycosin on breast cancer cells.Methods: To cultivate MCF-7 and T-47 D breast cancer cell lines,MCF-7 and T-47 D cells were treated with different concentrations of calycosin for 24 h,48 h,72 h,the proliferation of the cells treated with calycosin was tested by MTT assay.Apoptosis in MCF-7 cells was examined by Hoechst33258.To measure intracellular RCAS1 factor gene expression of calycosin in different concentrations separately applied to a certain number of MCF-7 cell and T-47 D breast cancer cell by Fluorescence quantitative PCR detection method,detect the expression of Foxp3,RCAS1 and VEGF protein factor levels in MCF-7 cell and T-47 D cells by using protein immunoblot(Western Blot)method.Results: Calycosin promoted apoptosis of MCF-7 and T-47 D cells with the increasing concentrations by MTT.According to the results,we select some different concentration for next experiments.Through the experiment of Hoechst 33258,promoting breast cancer cell apoptosis with the drug concentration increasingly..From the cells of gene and protein expression level,calycosin respectively inhibition of cancer cell for further research.Furthermore,the high concentrations of calycosin increased the expression of RCAS1 in breast cancer.With the high concentration in breast cancer cells,down-regulated protein levels of Foxp3 and VEGF,up-regulated the protein of RCAS1.Conclusion: Phytoestrogens calycosin down-regulate the protein levels of Foxp3 and VEGF,up-regulate the mRNA and protein levels of RCAS1 to inhibit proliferation and induce apoptosis of MCF-7 and T-47 D cells.