The Effect Of Follicle-stimulating Hormone On The Expression Of Notch1/Hey In The Ovarian Cancer Cells

At present,ovarian cancer is one of the three gynecological malignancies. Ovarian cancer is the most common histopathological type accounting for 85%-90% of all ovarian malignancies.Despite continuous improvement in surgical modalities, chemotherapeutic and radiotherapy over the past several years,the 5-years survival rate for ovarian cancer with the most poor prognosis of all gynecological malignancies is about 30%-40%,which has not improve significantly.Clinically,ovarian cancer is often found in postmenopausal women with high serum follicle stimulating hormone(FSH) and luteinizing hormone(LH) level.The risk for women who have many pregnancies,oral contraceptives,hormone replacement therapy is low,while their serum FSH and LH concentration is also low. In the past 20 years,previous studies have proven that FSH plays an important role in ovarian cancinogenesis.Recent studies showed that Notchl signaling pathway relates to the development of cancer,including carcinogenesis,invasion and migration.But little is known about the relationship between FSH and Notchl signaling pathway in ovarian cancer ceils.The result of our experiments suggests that FSH may effect Notchl signaling pathway activation in ovarian cancer cells.To better understand the relationship between different concentration of FSH and the activation of Notchl signaling pathway,different action time of FSH and the activation of Notchl signaling pathway,different concentration of FSH and the expression of Notchl signaling pathway's target gene,we could provide a new stratery for further studying the carcinogenesis of ovarian cancer.These experiments were devided into 2 parts,as follows:①the relationship between FSH and Notchl signaling pathway of ovarian cancer cells;②The effect of different concentration of FSH on the expression of Notchl signaling pathway's target gene in ovarian cancer cellsSectionⅠ.The relationship between FSH and Notchl signaling pathway in ovarian cancer cells Objective(1) To investigate whether FSH can effect the expression of Notchl ICD in ovarian cacer cell line SKOV-3 and ES-2 in vitro.(2) To investigate whether different concentration and different action time of FSH can effect the expression of Notchl ICD in ovarian cacer cell line SKOV-3 and ES-2 in vitro.Methods(1) Cultured SKOV-3 and ES-2 all lines were treated by FSH with different concentration(0,40mIU/ml).SKOV-3 cultured were 48h,ES-2 were cultured 24 h.Cellular immunochemistry detects the expression of Notchl ICD in SKOV-3 and ES-2 which had been treated or not treated by FSH.(2) Cultured SKOV-3 were treated by FSH with different action time(0,12,24,36,48,72h) and different concentration(0,10,20,40,80mIU/ml).Cultured ES-2 were treated by different time(0,12,24,36,48h) and different concentration of FSH(0,10,20,40,80mIU/ml).Western blot was used to detect the expression of Notchl ICD in SKOV-3 and ES-2 which had been treated with different concentration and different action time.Result Notchl ICD could be more obviously express in the nuclear of ovarian cancer cell lines SKOV-3 and ES-2 which had been treated by FSH than those which had not been treated by FSH.The expression of Notchl ICD protein in ovarian cancer cell lines SKOV-3 and ES-2 increased with the increased concentration and time of FSH.Conclusion FSH may has effection on expression of Notchl ICD in ovarian cancer cells,and FSH may enhance the activation.The expression level of Notchl ICD had positive correlation with the increased concentration and time of FSH in ovarian cancer cells.SectionⅡ.The effection of FSH on the expression of Notchl signaling pathway's target gene in ovarian cancer cellsObjective To investigate whether different concentration of FSH could impact the express of Notchl signaling pathway's target gene HEY in ovarian cacer cellsMethods SKOV-3 and ES-2 were treated by FSH with different concentration(0, 10,20,40,80mIU/ml).SKOV-3 were cultured 48h,ES-2 cultured were 24 h.RT-PCR was used to detect the expression of HEY—1,HEY—2 and HEY—L mRNA in SKOV-3 and ES-2,which had been treated with different concentration.Result FSH could increase the mRNA expressions of HEY—1,HEY—2 and HEY—L in ovarian cancer cell lines SKOV-3 and ES-2,but there is no obvious dose effect between them.Conclusion FSH may increase the level of the expression of Notchl signaling pathway's target gene HEY—1,HEY—2 and HEY—L in ovarian cancer cells without dose dependent.