| Objective: Endometrial cancer is a common gynecological malignancy and its incidence increases year by year,which is highly associated with elevated estrogen levels,early menarche,late menopause,infertility,long-term tamoxifen treatment and other factors.Postmenopausal bleeding is the most common clinical manifestation,and the five-year survival rate is about 80%.At present,the cornerstone of the treatment of endometrial cancer is surgery,supplemented by radiotherapy and chemotherapy.In recent years,molecular targeted drugs have become a new therapeutic method and entered the preliminary stage of clinical application in recent years.It is the fundamental approach of treatment to clarify the basis of disease occurrence and development and explore the pathogenesis.ETS related gene ERG,a member of ETS transcription factor family,is abnormally expressed in a variety of tumors.TCGA database shows that ERG is low expression in endometrial carcinoma.Lnc RNA plays an important role in regulating gene expression,growth,differentiation,development and other functions.Studies confirmed that Lnc RNA A2M-AS1 is involved in the occurrence of pancreatic cancer and breast cancer and associated with poor survival rate.TCGA database shows the low expression of A2M-AS1 in endometrial carcinoma.JASPAR and NCBI databases suggest that ERG has binding sites in the promoter region of A2M-AS1.At present,the mechanism of ERG and A2M-AS1 in endometrial cancer has not been reported.In this study,we will explore the mechanism by which ERG regulates the transcription of A2M-AS1 and influences the biological behavior of endometrial cancer cells.Methods: 1.To verify the expression levels of ERG and A2M-AS1 in endometrial carcinoma: Collected 20 normal endometrial tissues and 45 endometrial cancer tissues.qRT-PCR was used to detect mRNA levels of ERG in tissues.Western Blot was used to detect the protein level.Collected 20 normal endometrial pathology sections and 60 endometrial carcinoma pathology section.Immunohistochemistry was used to verify the location in endometrium,then analyzed the relationship between the protein level and clinicopathological parameters.qRT-PCR was used to detect the mRNA level of A2M-AS1 in tissues and analyzed its relationship with clinicopathologic parameters.2.To explore the effects of ERG and A2M-AS1 on the biological behavior of endometrial cancer cells: Cultured Ishikawa and HEC-1A cells,overexpressed and knocked down ERG and A2M-AS1,overexpressed and knocked down A2M-AS1 after the overexpression of ERG.Edu kit was used to detect proliferation ability,Transwell assay was used to explore invasion ability,Scratch assay was used to explore migration ability,Apoptosis kit was used to detect the level of early apoptosis.To explore the influence of ERG regulating the transcription of A2M-AS1 on the biological behavior of endometrial cancer cells: overexpressed ERG in two cell lines,qRT-PCR was used to detected the expression changes of A2M-AS1.JASPAR and NCBI databases were used to find the binding sites of ERG and A2M-AS1 promoter region,and ChIP-PCR was used to verify the binding of ERG and A2M-AS1 promoter region.Western Blot was used to detect the effects of ERG and A2M-AS1 on key molecules of MAPK pathway and EMT-related proteins.Results: 1.mRNA and protein expression of ERG is decreased in endometrial carcinoma,and is localized in vascular endothelium,which is associated with age.The mRNA expression of A2M-AS1 is decreased in endometrial carcinoma,which is related to Lymph node metastasis.2.In Ishikawa and HEC-1A cell lines,overexpression of ERG and A2M-AS1 can inhibit the proliferation,invasion and migration of endometrial cancer cells and promote early apoptosis.Knockdown of ERG and A2M-AS1 can promote the proliferation,invasion and migration of endometrial cancer cells,and inhibit early apoptosis.Knocking down A2M-AS1 after overexpression of ERG can partially rescue the inhibitory effect of overexpression of ERG on the proliferation,invasion and migration of endometrial cancer and the promotion effect on apoptosis level.Overexpression of ERG and A2M-AS1 at the same time can significantly inhibit the proliferation,invasion and migration of endometrial cancer cells,and significantly promote early apoptosis.ERG can regulate the expression of A2M-AS1 and bind to the promoter region of A2M-AS1.After overexpression of ERG,EMT-related indicators-the expression of E-cadherin is increased,while the expression of N-cadherin and Vimentin are decreased.The expression of ERK and JNK in MAPK signaling pathway are decreased.After overexpression of ERG,knocking down A2M-AS1 can partially rescue the increased expression level of E-cadherin and the decreased expression level of N-cadherin,Vimentin,ERK and JNK.Conclusion: The low expression of ERG and A2M-AS1 in endometrial carcinoma is related to age and Lymph node metastasis.ERG and A2M-AS1 can inhibit endometrial cancer by inhibiting the proliferation,invasion and migration of endometrial cancer cells and promote early apoptosis.By binding to the promoter region of A2M-AS1,ERG reduces the transcription level,inhibits the cascade amplification of MAPK signaling pathway and the process of EMT,inhibits the occurrence and development of endometrial cancer. |
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