17 Beta Estradiol FOXM1 Mediated Effect On Ovarian Cancer Cell Proliferation

Epithelial Ovarian malignant tumor(Ovarian malignant epithelial tumors), derived from the germinal epithelial Ovarian surface, is the most common pathological type, Ovarian cancer mortality has for years been comfortably in gynecological tumors for years, 5 years survival rate in patients with only from 15% to 20%, the Ovarian cancer therapy is the standard solution in the tumor cells to destroy the loss on the basis of combination chemotherapy with platinum drugs. Although most patients are with ovarian cancer in postmenopausal, but there are still about 40% of growth period caused by surgical treatment in patients with artificial menopause, the present with the improvement of living standards with living environment worsening pollution, particularly in the incidence of ovarian cancer in younger trend, the removal of the highly malignant cancer with radiation and chemotherapy after surgery to treat again, adding to the body, the young growth period after surgery face menopause symptoms caused by lower estrogen than natural menopause women more strongly, make limited survival period after surgery patients quality of life, and further promote the ovarian malignant tumor cancer mortality.Although, there are clinical experimental treatment, according to the results of 17 beta estradiol as a sex hormone drug used in menopause syndrome can make good clinical effect, but because of its role in the safety of the ovarian hormone replacement therapy after surgery mechanism is unclear, MWS found that hormone replacement therapy was also associated with increasing risk of ovarian cancer. Whether it can be 17 beta estradiol routine as a auxiliary drug use in the growth period of ovarian cancer patients with postoperative clinical treatment has yet been reached consensus.In recent years, the study found that FOXM1 transcription factors exist in a variety of organizations, and high expression in a variety of tumor tissues and cell lines, including ovarian cancer, FOXM1 transcription factor in epithelial ovarian cancer played an important role in occurrence and development of literature which shows that FOXM1 is closely related to the ER alpha receptor pathway, FOXM1 related to estrogen may be associated with ovarian tumor,which is expected to become a new gene therapy targets.Objective: this research is dedicated to study the expression of FOXM1 high estrogen and ER alpha with Different expression of the influence of the effect on the proliferation of ovarian cancer cell line growth, the possible mechanism was discussed preliminarily.First, for this reason, this study applies the Western blot detection method to detect the ovarian cancer cell line A2780 and SKOV3 in the expression of ER alpha, FOXM1, in order to further explore the E2, FOXM1 and FOXM1 blocker ther mosphacta element sulfur chain in ovarian cancer cell line A2780 and SKOV3 in vitro growth and the role of good cell model for screening and the related mechanism. This study try to explore the possible mechanism of E2 induced ovarian cancer cell growth in vitro, hormone replacement therapy after surgery for ovarian cancer provides theoretical basis for the safety.Methods: With containing 10% fetal bovine serum(fetal bovine serum, FBS), 0.01%penicillin streptomycin and 25 mm HEPES RPMI1640 culture, at 37 ℃, inclusive of our 5% saturated humidity our fleet of constant temperature incubator in conventional cultivation of human ovarian cancer cell lines A2780, as well as the degree of human SKOV3 ovarian serous capsule gland cancer cells, the daily observed under inverted microscope cell density and cell growth situation, every two days in a fluid, about need to extend the once a week, selection of logarithmic phase cell experiment. Various operations must be operating in ultra clean Taichung.Inverted microscope1 Application of inverted microscope to observe A2780 cells and SKOV3 cell morphology and take photos.2 Protein imprinting method(Western blot method: 3 methyl thiazole blue azo armor(MTS) method: the ovarian cancer cell line(SK0V3, A2780) for different concentration gradient(10-5M, 10-7M, 10-9M, 10-10 M, 10-12 M 10- 14 M, 10-16M) estradiol role in different time(24h, 48 h, 72h), not to add in RPMI1640 medium containing 10% fetal bovine serum estradiol drugs for blank control group, in order to contain less than 0.1% alcohol concentration in RPMI1640 medium containing 10% fetal bovine serum as a solvent in the control group. MTS method to test the effects of estradiol on cell growth, select the optimal concentration and time of drug action.3 Protein imprinting method(Western blot method: to detect A2780 and SKOV3 cell lines in the ER, FOXM1 protein expression level.and after different time different concentration gradient of estradiol on A2780 and SKOV3 cell lines in the ER, FOXM1 protein expression level of influence.4 Trans well experiment: 2 to 10-10 MB, 10 to 12 m, in the role of 10-14 m of estradiol SK0V3 and A2780 cells, RPMI1640 culture medium containing 10% fetal bovine serum as blank control group.24 h after continuous culture to take out the Trans well Chambers, fixed with 95% ethanol, using crystal violet stain, under inverted microscope(200 x) micro poous membrane computing moves to Trans well cell is lower the number of cells and evaluate migration and invasion ability and take photos.Results:1 Human ovarian cancer cell lines A2780 cells and SKOV3 cells: inverted microscope to observe the cell A2780, SKOV3 ovarian cancer growth in good condition, A2780 cells form for polygon transparent, bedclothes good cells, cell oval, round, compact connection between cells, SKOV3 cells in paving stone pattern, with the passage of time, the cell contact closely, and the population is growing again, as the typical form of epithetlioid cells.2 Western blot detection of ER in the A2780 were negative expression(no expression), ERA positive expression in SKOV3 cells lines, FOXM1 are expressed in the A2780, SKOV3 cells.Given estrogen ACTS on the A2780 cells and SKOV3 cells respectively48 h and72h, two kinds of FOXM1 expression in cells increased with the extension of time.3 The MTS test E2 on each cells proliferation after 24 h, 48 h, 72 h ability, found that, compared with control group, the A2780 cell lines extended over time and dose concentration increases with increasing proliferation ability.And on the basis of given E2 after 24 h to join FOXM1 blocker blocker thermosphacta element sulfur chain jointly develop present proliferation decline after48 h.SKOV3 cell lines extended over time and dose concentration increased proliferation present a downward trend, use high or low concentration E2 and sulfur chain wire rhzomorph FOXM1 blocking agent on SKOV3 cell lines after treatment had no effect on expression of proliferation.E2 concentration for 10-5 M, 10-10 M, 10- 12 M, 10 ~ 14 M drug effect time of 12-24 h, SKOV3 cells is most sensitive to drugs, inhibition of cell proliferation has the strongest promote cell apoptosis;To increase the drug concentration or prolonged drug can’t improve the cell apoptosis rate.4 Scratch experiment and Transwell little room detection ability of tumor cell migration, the results show that compared with the control group, 10 to 9 m, in the role of 10-10 m concentration of E2 A2780 cells for 24 h after observation, according to the results of giving E2 stimulus set of scratches healing area significantly greater than the control group(49.32)Conclusion: FOXM1 in A2780 cells mediated by estrogen dependent on proliferation. As estrogen concentration gradient increased effect on A2780 cell proliferation, concentration dependence, proliferation effect mainly depends on the concentration of E2, as the extension of E2 concentration and action time, A2780 cell growth rate increased significantly, E2 in SKOV3 cells with inhibition of cell growth, inhibit growth rate with the increase of concentration of E2 and evidently with the extension of time. Are statistically significant differences(P<0.01). FOXM1 proliferation effect may not be mediated by ER. But FOXM1 increase estrogen stimulation A2780 and SKOV3 cells prolife ration. Therefore, FOXM1 can promote cell proliferation of hormone replacement therapy, but depending onthe type and characteristics of cells, different concentration of estrogen effect on cell proliferation, concentration dependence, Its proliferation role mainly in estrogen concentration is quite high proliferation effect is not obvious, can choose closer to the physiological concentration when estrogen replacement therapy dose or slightly below the physiological dose concentration of E2.FOXM1 specific mechanism is not clear, need further research.