Effect Of Fructus Sophorae And Its Transformation Products On Proliferation And Apoptosis In Breast Cancer Cell Lines

The prevention and cure breast cancer effects of chinese herbs were more and more thought highly of now, especially the isoflavone which had been paid most close attention to. Sophora japonica L. is rich in isoflavone phytoestrogen, such as sophoricoside, genistin, genistein and so on, which are the effective component of anti-cancer effect. This paper mainly study on the effects of Fructus sophorae and its transformation products on breast cancer cell lines human breast cancer cells MCF-7 and MDA-MB-231 about growth inhibition, cell apoptosis induction and potential mechanisms. It provides theory evidence for the prevention and cure of breast cancer.Methods: Exponential phase human breast cancer cell line MCF-7(ER+)and MDA-MB-231(ER-)were treated by Fructus sophorae and its various transformation products in vitro, the survival rate of the cells were measured by a tetrazolium-based colorimetric assay(MTT)in different time. The drugs include Fructus sophorae, its transformation products, crude isoflavone, crude polysaccharides, purified genistein and isoprunetin. To study the effects of genistein and isoprunetin on MCF-7 cells, MTT assay was applied for cell proliferation detection, while Flow Cytometry(FCM) for cell cycle analysis, western blotting detection, the cell morphous observation and DNA ladder assay for cell apoptosis.First, studying the effect of Fructus sophorae and its transformation products on the human breast cancer cells show that Fructus sophorae and its transformation products (2 d) had no influence on the proliferation of MCF-7, while transformation products (16 d) could markedly inhibit the proliferation of MCF-7 at 72 h; Fructus sophorae and its transformation products promoted the growth of MDA-MB-231 by time-dependent manner.Then, studying the effect of Fructus sophorae and its transformation products (crude isoflavone, crude polysaccharides) on the human breast cancer cells show that the crude crude isoflavone (2 d,16 d) could markedly inhibit the proliferation of MCF-7; crude isoflavone (0 d,2 d,16 d) evidently promoted the growth of MDA-MB-231 by time-dependent manner. Moreover, crude polysaccharides had no influence on the proliferation of MCF-7, while evidently promoted the growth of MDA-MB-231.Finally, this paper had studied the effect of purified genistein and isoprunetin on the human breast cancer cells. The results showed that genistein and isoprunetin evidently inhibit the proliferation of MCF-7, while had no influence on the proliferation of MDA-MB-231. The typical morphological changes of apoptosis MCF-7 cells in isoprunetin group had been observed. Agarose electrophoresis could not detect cracked ladder-shaped DNA. Western blotting showed that the expression level of caspase-3 and PARP in MCF-7 was significantly increased by the treatment of genistein/isoprunetin; while no changes of PARP expression in MDA-MB-231 by long time treatment. There was an apoptotic peak in MCF-7 cells after incubating with isoprunetin for 72 h by flow cytometry. The cell apoptotic ratio was 12.1 %. Flow cytometry showed that the cell cycle might be blocked in S phase by genistein and G1-S phase by isoprunetin.Genistein and isoprunetin from Fructus sophorae biotransformation by Schizophyllum commune possessed certain anti-proliferative and pro-apoptotic effects on breast cancer cells. Initial investigation on molecule mechanisms indicated that these substance influence the activity of caspase-3.