The Regulation Of ARID1A In Endometrial Cancer Ishikawa Cells Can Be Caused By Activation Or Blocking Of ER

Objective: Endometrial cancer is one of the most common malignant tumor in female reproductive system,which has a rising morbidity and mortality in our country,and younger,damaging seriously to the women’s health.There are many factors associating with the occurrence and development of endometrial cancer,such as long-time estrogen stimulation without progesterone antagonist and tumor-suppressor gene inactivation.Our earlier research of this topic found that TAM,similar to estrogen agonists,can stimulate endometrial cells ER,leading to endometrial lesions, even endometrial carcinoma.After treated with TAM or E2,the expressions of ARID1 A in three types of differentiation of endometrial cancer cells were significantly lower than the untreated, and G0/G1 phase of cell shortened, while S phase lengthened.ARID1A(AT-rich interacting domain containing protein),as one of the reorganization of chromatin complex, is one of the newly discovered tumor suppressor genes. Its low expression is associated with a variety of tumorigenesis.Expression of ARID1 A in poorly differentiated uterine carcinoma and clear cell carcinoma decreased obviously.Zeng et al. found in glioma cells overexpression of ARID1 A downgraded p-Akt and p S6 K,prompting that ARID1 A regulats cell proliferation through PI3 K /Akt pathway.In ovarian clear cell carcinoma, the loss of ARID1 A were positively correlated with PIK3 CA mutations.Lowexpression of ARID1 A in endometrial carcinoma was relevant to mutation and activation of PI3 K pathway.ARID1 A can interact with PI3 K pathway, influencing the phosphorylation of Akt, so as to control the proliferation of tumor cells.Our preliminary experiment of this topic proves that ARID1 A gene in endometrial carcinoma were positively correlated with ER expression.PI3K/Akt signaling pathway is one of the important signal transduction pathways in cells, involved in regulating cell proliferation, apoptosis, and differentiation.Recent studies have shown that PI3K/Akt pathway activity is unusual in many tumors, which can cause not only cell’s malignand transformation but also the migration and adhesion of tumor cells and tumor angiogenesis.Research has shown that combining with the ER,estrogen can activat PI3K/Akt signal pathway of endometrial cancer cells interacting with some active factors, resulting in corresponding changes in the cell.How does loss of ARID1 A lead to the occurrence of endometrial cancer?There is no research at home and abroad.ARID1 A and ER all work through PI3K/Akt pathway,but whether there is a direct relationship or whether the relationship is through PI3K/Akt pathway is unclear.This experiment is aimed at comparing the chang of ARID1 A and ER in Ishikawa treated with estrogen receptor activator(TAM) or estrogen receptor blocker(ICI182780), thus to explore ARID1A’s role in occurrence and development of endometrial cancer and whether ARID1 A influences the PI3K/Akt signaling pathway by the change of ER,in order to offer preliminary theoretical basis for the pathogenesis of molecular biology in endometrial carcinoma.Methods:1 cell culture:The Ishikawa cells were cultured in untrient solution of 1640,including 10% fetal calf serum,at 37℃,5%CO2 saturated humidity incubator.The cells were digested by 0.25% trypsinase and then subcultured.Cells adhered to the botton in monolayer and were chosen to be used in the test at logarithmic growth phase.2 The expressions of ARID1 A,ERα,ERβ,PR,P53,PIK3 CA,Akt m RNA in Ishikawa treated with different concentrations of TAM or ICI182780 at different time respectively were detected by q RT-PCR.3 The expression of BAF250 a,ERα,ERβ,PR,P53,PIK3 CA,p-Akt protein in Ishikawa treated with different concentrations of TAM or ICI182780 at different time respectively were detected by Western-blot.4 The cell cycle and apoptosis of Ishikawa treated with different concentrations of TAM or ICI182780 at different time respectively were detected by flow cytometric method(FCM).Results:1 Treated by TAM, the expressions of ARID1 A, ERα, ERβ m RNA and protein reduced with the increase of drug concentration and the extension of drug processing time(P<0.05),However,the expressions of PR,P53,PIK3 CA, Akt m RNA and protein increased with the increase of drug concentration and the extension of drug processing time(P<0.05);Treated by ICI182780, the expressions of ARID1 A, ERα, ERβ m RNA and protein increased with the increase of drug concentration and the extension of drug processing time(P <0.05), However, the expressions of PR,P53,PIK3 CA,Akt m RNA and protein reduced with the increase of drug concentration and the extension of drug processing time(P <0.05).2 Treated with different concentrations of TAM at different time, G0/G1 phase of Ishikawa gradually shortened, while S phase lengthened with the increase of concentration and time(P<0.05),the change of apoptosis rate is not obvious.Treated with different concentrations of ICI182780 at different time,the apoptosis rate of Ishikawa gradually increased with the increase of concentration and time(P<0.05),the change of cell cycle is not obvious.Conclusions:After ER is activated or blocked,the expression of ARID1 A decreases or increases, which leads to the change of PI3K/Akt pathway and cell cycle.The occurrence of endometrial cancer may be related to the activation of ER, and then the inhibition of ARID1 A function and activation of Akt.