Synergy And Crosstalk Between Estrogen And Insulin On The Growth Of Endometrial Carcinoma

Endometrial carcinoma is one of the three gynecological malignancies,and the carcinogenesis mechanism is not clear yet. It is reported that the incidence of endometrial carcinoma was significantly increased. Therefore, it is particularly important to study the etiology and pathogenesis about EC.The risk factors of Endometrial carcinoma include obesity, diabetes, hypertension, long-term estrogen stimulating without progesterone.The common pathophysiological basis of these factors is insulin resistance and hyperinsulinemia. As a hormone, estrogen not only regulates the normal body growth and maintain female characteristic, but also can be used as a growth factor that promotes endometrial proliferation and the malignant transformation. Insulin can also regulate the growth and differentiation of cells through its downstream signaling pathways. As endometrial cancer risk factors, high estrogen and high insulin promote the malignant transformation of endometrial cells and they induce proliferation of endometrial carcinoma together. But how do the estrogen, insulin and their signal transduction interact in endometrial carcinoma? This study will use endometrial cancer cell line to discuss the synergy and crosstalk between estrogen and insulin on the growth of endometrial carcinoma cell line.SectionⅠThe expression of estrogen receptorαand insulin receptorβin endometrial carcinoma cell linesObjective:To investigate the expression of ER-a and INSR-βin endometrial carcinoma cell lines.Methods:The expression of ER-a and INSR-βin endometrial carcinoma cell lines were detected by RT-PCR and Western blot.These cell lines include AN3CA, Ishikawa, HEC1-A, HEC1B,KLE, ECC-1.Results:The six endometrial carcinoma cell lines were all existing the expression of ER-αand INSR-β.The expression of ER-a and INSR-βin Ishikawa and ECC-1 cell lines was more than others.Conclusion:The six endometrial carcinoma cell lines were all existing the expression of ER-a and INSR-P, and had the basis of estrogen and insulin signaling pathways. SectionⅡMitogenic effects induced by 17βEstrodial in endometrial carcinoma cell lineObjective:To explore mitogenic effects induced by 17-βEstrodial in endometrial carcinoma cell line.Methods:The mitogenic effects of different concentration of 17-βestradiol in endometrial carcinoma cell line were detected by MTT at different time. Experimental group:control, and 10-10 M 17-βestradiol,10-8 M 17-βestradiol,10-6 M 17-βestradiol, 10-4 M 17-βestradiol.Results:The effects of 17-βestradiol on Ishikawa cell at different time have statistically significant (F=25.117, P=0.000). Besides the concentrations of 0 M and 10-10M, the effects of various concentrations of 17-βestradiol have statistically significant (P<0.05). At the time of 72 hours, the effects reached the peak and then dereased.Conclusion:17-βestradiol could promote endometrial cell proliferation and dependent on the concentration and stimulating time.SectionⅢsynergy and crosstalk between estrogen and insulin on the growth of endometrial carcinoma cell lineObjective:To investigate synergy and crosstalk between estrogen and insulin on the growth of endometrial carcinoma cell lineMethods:The SiRNA of ER-αand INSR-βwere designed and synthetized.Screen effective sequence of ER-αand INSR-β. No-Serum hunger Ishikawa cells were divided into different groups.①To investigate the synergitic effects of insulin and 17-βestradiol, gouped:control,10-6M insulin treatment group,10-8M 17βestradiol treatment group,10-6M insulin plus 10-8M 17βestradiol treatment group;②To investigate the crosstalk effects of insulin and 17-βestradiol:ER-a interference groups, INSR-βinterference groups,co-transfection group, negative control.Each group was divided into control,17-βestradiol treatment group, insulin treatment group,17-βestradiol plus insulin treatment groups. the effects of cell proliferation were detected by MTT at different time.Results:The mitogenic effect of insulin plus 17βestradiol group were much more than insulin or 17-βestradiol group.After silencing ER-α, the effect of 17-βestrodiol was inhibited obviously.After silencing INSR-P, the effect 17-βestrodiol and insulin was inhibited obviously. After co-transfection, the effect of 17-βestrodiol and/or insulin were inhibited obviously.Conclusion:There are synergy and crosstalk between estrogen and insulin on the growth of endometrial carcinoma cell line.Above all, endometrial carcinoma cell line were existing the expression of ER-αand INSR-β, and have the basis of estrogen and insulin signaling pathways. Estrogen can promote endometrial proliferation. There are synergy and crosstalk between estrogen and insulin on the growth of endometrial carcinoma cell line.