Study On Apoptosis Of Breast Cancer Cells By Korean Soybean Paste Isoflavone Extract

Korean soybean paste is a traditional food which the Korean people particularly likes.Because of its unique flavor after fermentation,it is accepted by more and more people.After fermentation,not only the flavor changes,but also the bioactive substances inside.The main ingredient in the soybean paste is soy isoflavone,which have various effects such as anti-oxidation and anti-tumor,as well as estrogenic effects.Breast cancer is a disease with high incidence and mortality in women today.Combined with the efficacy of soy isoflavone,we have studied the effect of the isoflavone extract of Korean soybean paste on breast cancer cells on apoptosis.In order to study the changes of Korean soybean paste with fermentation,the main active components and the effect on the apoptosis of breast cancer cells,this experiment used laboratory homemade Korean soybean paste.It was sampled in different fermentatioa periods(0,1,3,6,9,12 months).And their components were determined by High Performance Liquid Chromatography(HPLC).Choose the best storage period of Korean soybean paste for extraction.The effect of IEOKSP on proliferation of normal breast cells,MCF-7 and MDA-MB-231 breast cancer cells was determined by CCK-8 assay.The healing degree was observed through the scratch healing experiment.Flow cytometry was used to detect the apoptosis of MDA-MB-231 and MCF-7 breast cancer cells treated with IEOKSP at different concentrations,as well as the cycle changes.The expression of caspase-3 mRNA was detected by RT-PCR.The expressions of Bax,Bcl-2,Survivin,caspase-3 and cleaved caspase-3 were detected by Western Blot.Finally,the energy metabolism of MDA-MB-231 and MCF-7 breast cancer cells treated with IEOKSP was detected by Sea House cell energy analyzer.The results showed that the isoflavone components in the extract of Korean soybean paste changed with the fermentation and the extension of storage time.The contents of daidzein and genistein increased gradually with the extension of storage time,while the contents of daidzein decreased gradually.IEOKSP can inhibit the cell activity of MDA-MB-231 and MCF-7 breast cancer cells in a concentration-time dependent manner,and can effectively inhibit cell migration,block cell G1 phase,and induce apoptosis.IEOKSP can increase the expression of caspase-3 mRNA in MDA-MB-231 and MCF-7 breast cancer cells,up-regulate the expression of Bax,caspase-3,cleaved caspase-3 protein,and down-regulate the expression of Bcl-2,survivin protein,leading to apoptosis.IEOKSP can reduce the oxygen consumption rate of breast cancer cells through mitochondrial pathway.It can be concluded that the main functional components in IEOKSP are gradually increased with the prolongation of storage time,and can inhibit the proliferation of breast cancer cells by regulating Bax,caspase-3,cleaved-caspase-3,Be 1-2,survivin protein expression,blocks cell cycle,and prevents and adjuvant treatment of breast cancer.