MiRNA-302a-5p/miR-367-3p Affects The Malignant Biological Behavior Of Endometrial Cancer Cells By Targeted Regulation Of HMGA2

Objective:The incidence of endometrial cancer,one of the malignant tumors of the female reproductive system,has increased year by year,and has now become a common female malignant tumor in developed countries.Treatment for endometrial cancer includes surgery,supplemented by radiation,chemotherapy,and hormones or other medications.However,for patients with advanced metastatic or recurrent endometrial cancer,treatment failure remains high due to the loss of surgical opportunity.With the elucidation of the molecular mechanism of endometrial cancer,more and more molecular targeted drugs are being tested in clinical applications,and some have shown beneficial effects,the clinical application of EFGFR2 and mTOR signaling pathway inhibitors has achieved some success.Obtained preliminary clinical application and achieved certain effects.Therefore,the underlying mechanisms of endometrial cancer development,especially those involved in tumor metastasis,have become the basis for the development of molecular targeted therapy for endometrial cancer,which is essential for the treatment of such cancers.The high mobility group protein A(HMGA)family is a small class of non-histone chromosomal proteins characterized by an AT-hook domain and an acidic end,including four members of HMGA1a,HMGA1b,HMGA1c and HMGA2.Among them,the coding gene of HMGA2 is located in the cluster region of chromosome breakpoints of many tumor cells,and its abnormal expression is related to tumors,and its biological behavior is similar to oncogene.High mobility histone A2(HMGA2)has become a candidate biomarker.It is worth noting that the high mobility group AT-hook 2(HMGA2)has been considered as a driver of tumor metastasis.MicroRNAs(miRNAs)are a class of non-coding small RNAs with regulatory functions that are about 19-25 nucleotides in length.It mainly participates in the regulation of apoptosis and proliferation by completely or incompletely pairing the seed region with the 3' untranslation region(3'-UTR)of the target gene mRNA,degrading the mRNA of the target gene or inhibiting its translation.And a variety of physiological and pathological processes such as differentiation,is a powerful post-transcriptional regulator of HMGA2.The aim of this study was to investigate the malignant biological behavior of non-coding RNAs in the regulation of endometrial cancer cell lines,and to further analyze the possible molecular mechanisms:miRNA-302/367 regulates the malignant biological behavior of endometrial cancer cell lines by targeting HMGA2.Results:Real-time-PCR was used to detect the expression level of HMGA2 in human endometrial carcinoma tissues,and to analyze the relationship between HMGA2 expression and clinicopathological parameters of patients with endometrial cancer.Western blotting and immunohistochemistry were used to detect HMGA2 in human endometrium.Expression levels in cancerous tissues.The expression levels of miR-302a-5p and miR-367-3p in human endometrial carcinoma tissues were detected by Real-time-PCR,and analyze the relationship HMGA2 expression and clinical pathological parameters in endometrial cancer.The binding sites of miR-302a-5p and miR-367-3p of HMGA2 mRNA were identified by bioinformatics prediction software,and HMGA2 wild-type and mutant luciferase vectors were constructed,which were combined with miR-302a-5p and miR.-367-3p was co-transfected with 293T cells,and the binding and binding sites of HMGA2 to miR-302a-5p and miR-367-3p were detected by dual luciferase reporter assay.The Ishikawa and HEC-1A cell lines of HMGA2 overexpressing and knocking down endometrial carcinoma were constructed,the real-time PCR assay was used to verified and the transfection effect.The CCK-8and edu assay is used to detect the overexpression and knockdown of HMGA2 in both cells;scratch assay to detect changes in cell migration capacity after the same treatment;Transwell assay to detect cell invasion after treatment with the same method Ability to change.Flow cytometry was used to detect changes in cell cycle after intervention of HMGA2 expression;flow cytometry was used to detect apoptosis in the same treatment;miR-302a-5p/miR-367-3p was constructed by cell transfection In overexpressed and silenced cell lines,CCK-8 assay detects changes in miR-302a-5p/miR-367-3p overexpression and cell proliferation in both cells;the EdU assay also detects miR-302a-5p/miR-367-3p overexpression level and proliferative capacity of Ishikawa and HEC-1 A cells after intervention;scratch assay for miR-302a-5p/miR-367-3p overexpression and cell migration after intervention Capacity changes;Transwell assay detects changes in miR-302a-5p/miR-367-3p overexpression and cell invasiveness after intervention.Flow cytometry was used to detect the changes of cell cycle after miR-302a-5p/miR-367-3p expression;flow cytometry was used to detect apoptosis after the same treatment;Western Blot method was used to detect N-cadherin,expression changes of E-cadherin protein.The effects of miR-302a-5p/miR-367-3p and HMGA2 on the growth of xenografts in nude mice were tested in nude mice xenografts.The in vitro and in vivo assays were used to study the passage of miR-302a-5p and miR-367-3p.Targeted regulation of the effects of HMGA2 on the malignant biological behavior of endometrial cancer cells.Conclusion:1.HMGA2 is highly expressed in endometrial carcinoma tissues,miR-302a-5p/367-3p is underexpressed in endometrial carcinoma tissues,miR-302a-5p and miR-367-3p expression levels and FIGO stage and lymph node metastasis closely related.HMGA2 and miR-302a-5p/367-3p are negatively correlated in endometrial cancer tissues.2.HMGA2 targets binding to miR-302a-5p/367-3p.3.miR-302a-5p/367-3p can inhibit the malignant biological behavior of endometrial cancer cells by negatively regulating HMGA2.