The Effects Of Genistein On Primary Cultured Ectopic Endometrial Cells In Vitro And Its Mechanism

Objective:To explore the inhibitory effect of Genistein(GEN)on mouse endometriotic cell cultured in vitro and its mechanism of apoptosis.Method:30 female Balb/c mice of 6-7 weeks old weighing 20g were selected to prepare an animal model of endometriosis in mice.Tissues collected from the uterus of normal mice,the uterus of endometriosis modeled mice and the implanted graft,were cut into pieces,degraded,incubated and finally filtered to obtained cells,primary culture were performed in DMEM medium and the morphology and growth pattern of the cells were observed under electron microscope.The normal endometrial cell,EMs eutpoic cells and EMs ectopic cells were then treated with different dose of GEN(0.1 ?M,1 ?m,10 ?M)for 24 hours.The effect of GEN on the cells were observed by using MTT assay and the apoptosis rate of each group was observed by Annexin-V FITC/PI staining.The expression of Caspase-8 and NF-?B in EMs ectopic endometrium before and after the treatment with high dose(10?M)GEN was detected by Semi quantitative PCR.Result:1)Normal endometrial cells,EMs eutopic endometrial cells and EMs ectopic endometrial cells was successfully cultured in vitro.Under electron microscope,it was observed that the stromal cells of EMs ectopic cells were smaller than those of stromal cells of EMs eutopic cells and normal endometrial cells,and the surface was irregular with more microvilli.2)The growth rate of normal endometrial cells and EMs eutopic cells is slower than that of EMs ectopic cells.Annexin V/PI staining,showed that the apoptosis rates of the three types of cells were 34.5%,32.I%,and 20.6%respectively,indicating a gradual decreasing trend.3)The 24 hour growth rate of EMs ectopic endometrial cells after treated with different concentrations of GEN was 99.74�3.21%,94.00�2.51 and 71.58�4.42%,respectively.The high dose of GEN inhibited the growth of ectopic endometrial cells of EMs(p<0.05).4)The number of apoptotic cells and dead cells after high-dose GEN treatment in EMs ectopic endometrial cells were found to be increased than before the treatment and the amount of NF-kB mRNA showed a decreasing trend,where the amount of Caspase-8 mRNA increases.Conclusion:1)High concentration of GEN can inhibit the proliferation of ectopic endometrial cell cultured in vitro and at the same time promotes apoptosis.2)GEN may exert its inhibitory effect on ectopic endometrial cell through the down regulation of NF-?B and up-regulation of Caspase-8 expressions.