Expression Of SOX17 And CyclinD1 In Endometrial Carcinoma And The Effect Of 5-AZA On Its Expression

Background and PurposeEndometrial cancer(EC)is a common female gynecological malignancy tumor.In recent years,the incidence rate is on the rise,especially the incidence of young patients.The pathogenesis of endometrial cancer is not clear,epigenetic changes in the development and progression of the tumor plays an important role.For the time being,the role of SOX17 in the development of endometrial cancer is unclear.The aim of this study was to investigate the relationship between SOX17 and endometrial cancer.SOX17 is an important antagonist of the classical Wnt signaling pathway.It regulates Wnt / ?-catenin signaling by inhibiting ?-catenin / TCF-dependent transcription and cancer cell proliferation and colony formation.A number of studies have shown that high methylation of the SOX17 promoter can reduce the expression of SOX17 in tissues,and finally leading to the proliferation of cancer cells.Further studies have shown that SOX17 inhibits tumor mechanisms that may be associated with its ability to block cells in G1 / S phase,and induce apoptosis and / or regulate the angiogenesis of tumor cells.It is concluded that SOX17 may play a role in the process of human cancer,but its potential role in human endometrial cancer has not been elucidated.Human tumor is a cell cycle disease.As a positive regulator of the cell cycle,cyclin D1 promotes the proliferation of tumor cells.Methyltransferase inhibitor 5-azacytidine(5-AZA)is able to re-express the over-methylated gene by demethylation to restore its anti-cancer function.The aim of this study was to investigate the mRNA expression of SOX17 and CyclinD1 in EC tissues.Then using different concentrations of 5-AZA to interfereEC cells and detect the different mRNA expression of SOX17 and CyclinD1 aiming to detect the expression of SOX17,CyclinD1 and ?-catenin and study the effect of 5-AZA on the expression of SOX17 and CyclinD1 in EC cell lines.Material and Methods 1Experimental material 1.1 Cell Source:Human endometrioid adenocarcinoma cell line HEC1 A,donated by the key laboratory of the First Affiliated Hospital of Zhengzhou University.1.2 Tissue specimen source and case data:Collected Tissue specimen source in July 2014 ~ December 2016 in Zhengzhou University,the Second Affiliated Hospital of hysterectomy endometrial tissue specimens,all the intraoperative specimens were immediately preservated at-80?.The specimens consisted of 30 cases of endometrioid adenocarcinoma and 10 cases of normal proliferative endometrial tissues.All specimens were confirmed by the pathology department of the Second Affiliated Hospital of Zhengzhou University.2Methods 2.1 Experimental group:the collected specimens were divided into two groups,30 cases of endometrioid adenocarcinoma group,10 cases of normal proliferative endometrial group(due to cervical intraepithelial neoplasia surgery to remove the uterus endometrial tissue).Patients in the two groups did not accepted radiotherapy,chemotherapy,hormone immunotherapy and other anti-tumor therapy.Those patients were excluded with other parts of the tumor.Allthe tissue samples were observed by the pathologist physician microscopic diagnosis.2.2 The expression of SOX17,?-catenin and CyclinD1 genes in EC tissues and endometrium tissues in normal:Using real-time fluorescence quantitative PCR detect the expression of SOX17,?-catenin and CyclinD1 in 30 cases of endometrioid adenocarcinoma and 10 cases of normal proliferative endometrium.2.3 The impact of 5-AZA on the proliferation of endometrioid adenocarcinoma cells:Different concentrations of 5-AZA treatedwith HEC1 A.MTT assay was used to detectcell's growth inhibition.The mRNA expression of HEC1 A was detected by real-time quantitative PCR.3.Statistical analysisThe data was analyzed by SPSS 23.0 software and represented in the form of (?).Differences among groups were analyzed by using Independent Sample t Test,ANOVA and Pearson correlation analysis,with P<0.05 as statistically significant,? = 0.05 for the test level.Results 1.The different mRNA expression of SOX17,?-catenin and CyclinD1 in Normal Proliferative Period Endometrial Tissue and Endometrial Carcinoma tissueIn this study,we found that the expression of SOX17 gene was down-regulated in EC tissues.Meanwhile,the expression of ?-catenin and CyclinD1 were up-regulated in EC tissues.Compared with normal endometrial tissue,the expression of SOX17 gene was down-regulated in EC tissues and the result has statistically significant(P <0.05).The relative expression of SOX17,?-catenin and CyclinD1 mRNA was significantly different in histological grading,lymph node metastasis and FIGO staging(P<0.05).The relative expression of ?-catenin and CyclinD1 mRNA in the depth of myometrial invasion was statistically significant(P <0.05).2.Effects of 5-AZA on the proliferation of EC cells3.Correlation analysis of SOX17 and CyclinD1 genes in endometrial carcinoma5-AZA has been shown to be an effective anticancer drug,but its specific mechanism is unclear.In this study,we investigated the effect of 5-AZA on Wnt-?-catenin signaling in endometrial carcinoma cells.MTT assays were used to detect the inhibitory effect of 5-AZA on the growth of HEC1 A cells at different concentrations of 24 h,48h and 72 h.There was a significant difference between virous concentrations of the inhibitory effect of 5-AZA on the growth of HEC1 A cells(P<0.05),and its growth inhibition effect was significant(IC50 = 12.033).When added different concentrations of 5-AZA to HEC1 A for 24 h,48h,72 h,with the increase of interference time,the growth inhibition effect was enhanced.The inhibitory effect of HEC1 A cells growth on different time was statistically significant(P <0.05).In this study,the relative mRNA expression of CyclinD1,?-catenin and SOX17 in 30 cases of EC tissues was analyzed by pearson correlation analysis.The expression of ?-catenin and CyclinD1 has showed positive correlation in EC tissues(r = 0.863,P< 0.001);The expression of SOX17 was negatively correlated with CyclinD1 and ?-catenin in EC tissues(r =-0.353,P> 0.05;r =-0.463,P< 0.05).Conclusions 1.SOX17 gene may play an important role in the gene therapy of endometrioid adenocarcinoma.It can down-regulate the expression of CyclinD1 and ?-catenin geneand achieve thepotential tumor suppressor function.2.Demethylated drug 5-AZA can enhance the expression of SOX17 gene in endometrioid adenocarcinoma cell lines and inhibit the proliferation of endometrial cancer cells.In the future treatment of endometrial carcinoma,it caneffect on SOX17 to achieve its anti-cancer functionand become the Potential effective anti-cancer drugs.