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Objective: the different concentrations icaritin (Icaritin, ICT) used alone and in combination with estrogen application, MCF-7 breast cancer cell proliferation and inhibition, and analysis of the expression of the estrogen receptor ERα in the process and explore possible ways of ICT estrogen and anti-estrogen effect, to treat breast cancer may be looking for a surgery with the method provides a theoretical basis. Method: 1. Vitro culture of estrogen-dependent breast cancer cells MCF-7 cells with different concentrations of ICT (10-10, 10-9, 10-8, 10-7, 10-6, 5 × 10-6, 10-5) on different concentrations the E2 (10-13,10-12,10-11,10-10,10-9,10-8,10-7,10-6) and 10-9 E2 different concentrations ICT (10-7,10-6,5 × 10-6,10-5,5 × 10-5) role in MCF-7 cells after 6 days, the morphological changes of MCF-7 cells were observed under an inverted microscope. With different concentrations of ICT (10-10,10-9, 10-8,10-7, 10-6,5 × 10-6,10-5,5 × 10-5) with different concentrations of E2 ( 10-13,10-12,10-11,10-10,10-9,10-8,10-7,10-6) E2 different concentration of 10-9 ICT (10-8,10-7 , 10-6,5 × 10-6,10-5,5 × 10-5) role in MCF-7 cells four days, five days, six days after the MTT assay cell proliferation inhibition changes, and calculate the proliferation rate and inhibition rate. 3.10-9 E2 different concentration of ICT (10-7, 10-6,5 10-6,10-5) role in MCF-7 cells after 6 days, Hoechst33342 staining of apoptotic cells, and calculate the rate of apoptosis. 4. The icaritin estradiol combined with a luciferase reporter gene assay kit to detect ERα reporter gene active. 5.RT-PCR the detection icaritin and estrogen combination MCF-7 cells, estrogen receptor ERα expression changes. Results: 1. Estrogen alone deal with MCF-7 cells, the cell division phase transition, some of the cells into polygons, more and more, the number of cells with the increase in the concentration of estrogen E2 when E2 concentration reached 10-9 when cells from either the quantity or split-phase maximum. After the number of cells and the split-phase gradually become less. . Icaritin separate treatment of MCF-7 cells, the cell division phase transition, some of the cells into polygons, more and more, the number of cells with the increase in the concentration of estrogen ICT ICT reach 10-5 cell division maximum cell number and split phase after gradually become less. 3.10-9E2 concentration of ICT (10-7, 10-6,5 10-6,10-5) role in MCF-7 cells, cell division less, some cells became round, when the concentration of 10 - 5:00 cells floating and most obvious necrosis. 4.MTT colorimetric measurement result is displayed in the concentration of icaritin 10-6-5 × 10-5mol / l concentration range may be different ranges for MCF-7 cell proliferation, when the concentration of 10-5mol icaritin / l, the activity of promoting proliferation of the most obvious. Compared with estrogen E2 icaritin have weak estrogenic activity. 5 icaritin estradiol joint used in MCF-7 cells, MTT colorimetric measurement results show that when concentration 10-9mol / l of E2 concentrations greater than 10 (-9) mol / l of ICT joint used in MCF-7 cells, MCF-7 cell proliferation rate was significantly lower than the same concentration of E2 alone. 6.ICT alone can induce ERα reporter gene expression, When ICT and E2 in combination ICT on the E2-activated ERα reporter gene and inhibition. The 7. The icaritin used in MCF-7 cells with estradiol joint, RT-PCR test results showed that can inhibit ERα expression. Conclusion: 1. The icaritin within a certain concentration range can promote the proliferation of MCF-7, the estrogen-like effects. 2. The icaritin estradiol combined effects can be antagonistic former estrogen effect, inhibiting the latter promote MCF-7 cell proliferation. . Icaritin estrogen-like effects may occur through the estrogen receptor ERα selective estrogen receptor modulators (SERMs) role.
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