miR-7-5p、ABCC1在子宫内膜癌组织中的表达及对Ishikawa细胞增殖、EMT的影响

Expression of miR-7-5p and ABCC1 in endometrial carcinoma and their effects on Ishikawa cell proliferation and EMT

目的探讨微小RNA-7-5p(miR-7-5p)、ATP结合盒亚家族C成员1(ABCC1)在子宫内膜癌(EC)组织表达及其对EC细胞(Ishikawa)增殖及上皮间质转化(EMT)的影响。方法选取2018年6月至2021年12月期间在保定市妇幼保健院进行手术治疗的120例EC患者作为观察对象,手术取癌组织及癌旁组织,检测EC组织与癌旁组织miR-7-5p、ABCC1 mRNA及蛋白表达水平,分析miR-7-5p、ABCC1与EC临床病理特征的关系。将Ishikawa细胞随机分为Control组、miR-NC组、miR-7-5p mimics组、si-NC组,si-ABCC1组、miR-7-5p mimics+pcDNA组、miR-7-5p mimics+ABCC1组,RT-qPCR检测各组细胞miR-7-5p、ABCC1 mRNA表达水平,MTT法与流式细胞仪分别检测细胞增殖与凋亡,Western blot检测ABCC1、E-cadherin、α-catenin、Vimentin蛋白表达,双荧光素酶报告实验验证miR-7-5p与ABCC1的靶向关系。结果与癌旁组织比较,EC组织中miR-7-5p表达水平显著降低,ABCC1 mRNA及蛋白阳性表达率显著升高,相关性分析显示,miR-7-5p与ABCC1 mRNA水平呈负相关性(r=–0.580,P<0.05);miR-7-5p、ABCC1与EC分化程度、TNM分期及淋巴结转移有关(P<0.05);过表达miR-7-5p或抑制ABCC1表达可显著抑制Ishikawa细胞增殖,促进细胞凋亡,并上调E-cadherin、α-catenin表达,下调ABCC1、Vimentin蛋白表达水平;过表达ABCC1可部分逆转过表达miR-7-5p对Ishikawa细胞增殖及EMT的抑制作用;双荧光素酶报告实验证实miR-7-5p可靶向调控ABCC1表达。结论EC组织中miR-7-5p下调表达,而ABCC1上调表达,过表达miR-7-5p可通过抑制ABCC1表达抑制EC细胞增殖及EMT。

Objective To investigate the expression of microRNA-7-5p(miR-7-5p)and ATP-binding cassette subfamily C member 1(ABCC1)in endometrial cancer(EC)tissues and their effects on proliferation and epithelial-mesenchymal transition(EMT)of EC cells(Ishikawa).Methods A total of 120 EC patients who underwent surgical treatment in Baoding Maternal and Child Health Hospital from June 2018 to December 2021 were selected as observation objects,and the cancerous tissues and adjacent tissues were taken surgically,and the expression levels of miR-7-5p and ABCC1 mRNA and protein in EC tissues and adjacent tissues were detected,the relationship between miR-7-5p,ABCC1 and clinical pathological characteristics of EC was analyzed.Ishikawa cells were randomly divided into Control group,miR-NC group,miR-7-5p mimics group,si-NC group,si-ABCC1 group,miR-7-5p mimics+pcDNA group,and miR-7-5p mimics+ABCC1 group,the expression levels of miR-7-5p,ABCC1 mRNA in each group were detected by RT-qPCR method,the cell proliferation and apoptosis were detected by MTT assay and flow cytometry,respectively,the protein expressions of ABCC1,E-cadherin,α-catenin and Vimentin were detected by Western blot,the targeting relationship between miR-7-5p and ABCC1 was verified by double luciferase report experiment.Results Compared with the adjacent tissues,the expression level of miR-7-5p in EC tissues decreased obviously,and the ABCC1 mRNA expression and protein positive expression rate increased obviously,correlation analysis showed that there was a negative correlation between miR-7-5p and ABCC1 mRNA levels(r=–0.580,P<0.05).miR-7-5p and ABCC1 were related to EC differentiation,TNM stage and lymph node metastasis(P<0.05).Overexpression of miR-7-5p or inhibition of ABCC1 expression could significantly inhibit the proliferation of Ishikawa cells,promote cell apoptosis,up-regulate the expression of E-cadherin andα-catenin,and down-regulate the expression of ABCC1 and Vimentin.Overexpression of ABCC1 could partly reverse the inhibition of overexpression of miR-7-5p on Ishikawa cell proliferation and EMT.The double luciferase report experiment confirmed that miR-7-5p could target and regulate the expression of ABCC1.Conclusion The expression of miR-7-5p is down-regulated,while ABCC1 is up-regulated in EC tissues.Overexpression of miR-7-5p can inhibit EC cell proliferation and EMT by inhibiting ABCC1 expression.