Efforts Of Estrogen On The Expression Of EGF&EGFR Gene And Protein For Endometrial Cells

Objective:In this study, through the establishment of the reign of ectopic endometrial cells in primary culture model, ectopic&eutopic endometrium cells in primary culture morphology was observed, while the different dose&different estrogen stimulation of EGF&EGFR gene and protein expression changes in order to understand the relationship of endometriosis in EGF&EGFR expression with estrogen.Methods:The test specimens from Tianjin Medical University Second Hospital gynecological patients due to the EM surgical treatment of ectopic endometrial tissue as ectopic endometrium group (17cases), at the same time take the the uterine fibroids EM line the hysterectomy surgical treatment of endometrialeutopic endometrium (n=17), over the same period of hospitalization due to uterine fibroids underwent surgery in patients with endometrial for the control group (17cases), cell culture, the primary culture ectopic and eutopic endometriumThe cell morphology was observed. Cells in each group intervention methods:â‘ the cells in each group at different concentrations (10-4,10-6,10-8,10-10mmol/L) estrogen culture5d, to determine the optimal stimulation of estrogen concentration, collection of different concentrationsstimulation of the cells in each group and set aside. Were incubated30min and5d.â‘¡cells in each group to the optimum concentration of estrogen-containing medium, cells were collected and set aside. Trace enzyme reaction with tetrazolium salt (MTT) colorimetric method to detect the expression of the proliferation of cells in each group EGF mRNA and EGFR mRNA expression level of application of quantitative PCR detection by ELISA protein level detection of EGF and EGFR protein expression.Results:(1) The growth curves of vitro ectopic endometrium of endometriosis cells, eutopic endometrium cells and control cells are very close. Doubling time appeared at first3d,6d each group in the first cell proliferation are declining. Observed under inverted microscope, ectopic endometrial cells in endometrium and the control group of similar appearance characteristics, ectopic endometrial stromal cells form a slightly smaller endometrial stromal cells in control group, scattered grow, like a spindle.(2) Estrogen stimulation can improve the ectopic endometrial cells in endometrial cells and control cells proliferation capacity, and with the increase of concentration of estrogen stimulation, each cell proliferation ability are enhanced.(3) Ectopic and eutopic endometrial cells EGF, EGFR gene and protein expression in endometrial cells compared with the control group, both of which were increased expression, difference have statistical significance (P<0.05), and the EGF, EGF gene and protein expression with estrogen concentration dependence, groups of cells within the different concentration of estrogen stimulation between two comparative differences are statistically significant (P<0.05).(4) after the short-term and long-term estrogen stimulation, ectopic of EGF, EGFR in endometrium cells and gene and protein expression was increased endometrial cells, the difference had statistical significance (P<0.05), whereas estrogen comparing differences between long-term and short-term stimulus had no statistical significance (P>0.05).Conclusions:(1) Selecting eutopic and ectopic endometrial cells in primary culture model, is a reliable method, experimental results obtained with the reference value.(2) Estrogen can increase endometriosis endometrial cells EGF, EGFR gene and protein expression, which is proliferation of estrogen-related, and with the increased concentration of estrogen or estrogen to be short-acting and long-lasting stimulation, ectopic endometrial cells group EGF, EGFR gene and protein expression compared with eutopic endometrial cells and control groups were strengthened.(3) Ectopic endometrium and eutopic endometrial cells in cell groups of EGF, EGFR gene and protein expression and independent of estrogen stimulation duration.(4) Estrogen Endometriosis promoting proliferation of endometrial cells may be associated with enhanced expression of EGF and EGFR relevant.