子宫内膜癌中SFRP4和miR-421的表达与意义  

Expression and significance of SFRP4 and miR-421 in endometrial cancer

作　　者：高美 陈茜[1] 王颖[1] GAO Mei;CHEN Qian;WANG Ying(Department of Gynaecology and Obstetrics,Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710000,China)

机构地区：[1]西安交通大学第二附属医院妇产科,西安710000

出　　处：《山西医科大学学报》2023年第2期156-161,共6页Journal of Shanxi Medical University

基　　金：陕西省自然科学基础研究计划项目(2017JM8153)。

摘　　要：目的探讨分泌型卷曲相关蛋白4(SFRP4)在子宫内膜癌细胞系中的表达和miR-421对SFRP4表达的调控作用。方法分析基因表达综合数据库(GEO)中子宫内膜癌样本Microarray测序结果(GSE106191)寻找差异表达基因,对照组为数据库中同批次测序的良性增生样本。利用癌症基因组学数据分析平台(UCSC Xena)分析SFRP4表达对子宫内膜癌患者总生存期及肿瘤分级的临床意义。通过Targetscan软件找到可能与SFRP4结合的microRNAs。以子宫内膜癌细胞系(HEC-1A细胞)为实验对象,分为阴性对照组(miR-NC)、miR-421类似物组(miR-421 mimics)和miR-421抑制剂组(miR-421 inhibitor),分别转染阴性对照试剂、miR-421类似物和抑制剂,利用实时定量PCR和CCK-8法检测其对SFRP4 mRNA表达及细胞增殖能力的影响。为观察过表达SFRP4蛋白是否可回转上述改变,将HEC-1A细胞分为阴性对照组(miR-NC)、miR-421类似物组(miR-421 mimics)、miR-421类似物组+阴性质粒转染组(miR-421 mimics+vector)和miR-421类似物组+过表达质粒转染组(miR-421 mimics+pcDNA-SFRP4),分别转染阴性对照试剂、miR-421类似物、miR-421类似物和阴性质粒、miR-421类似物和SFRP4过表达质粒(pcDNA-SFRP4),利用CCK-8法检测其对HEC-1A细胞增殖能力的影响。结果Microarray测序结果发现SFRP4基因为差异表达基因,在子宫内膜癌中表达降低(P<0.0001)。SFRP4基因表达影响子宫内膜癌患者总生存期(P<0.01),但与临床分期无关(P>0.05)。利用Targetscan软件发现与SFRP4结合的microRNAs为miR-421。与阴性对照组相比,miR-421类似物组SFRP4 mRNA的表达水平降低,细胞增殖能力增强(均P<0.05);miR-421抑制剂组SFRP4 mRNA的表达水平增高,细胞增殖能力减弱(均P<0.05)。与miR-421类似物组+阴性质粒转染组相比,miR-421类似物组+过表达质粒转染组细胞增殖能力明显减弱(P<0.05)。结论SFRP4在子宫内膜癌中低表达,而miR-421通过靶向调控SFRP4的表达水平促进子宫内膜Objective To investigate the expression of secreted frizzled related protein 4(SFRP4)in endometrial cancer cell lines and the effect of miR-421 on SFRP4 expression.Methods Microarray sequencing results about endometrial carcinoma samples in gene expression omnibus(GEO)database(GSE106191)were analyzed to find out the differentially expressed genes,with benign hyperplasia samples in the same database as control.The relationships of SFRP4 expression with clinicopathological characteristics and overall survival were analyzed using Cancer Genomics Data Analysis Platform(UCSC Xena website).The microRNAs that may bind to SFRP4 were predicted using Targetscan tools.The endometrial cancer cell line HEC-1A cells were divided into negative control group(miR-NC),miR-421 mimics group(miR-421 mimics)and miR-421 inhibitor group(miR-421 inhibitor),and transfected with negative control reagents,miR-421 analogs and inhibitor,respectively.The effects of miR-421 on SFRP4 mRNA expression and cell proliferation were detected by real-time quantitative PCR and CCK-8 methods.To observe whether SFRP4 protein overexpression can reverse the above changes,HEC-1A cells were divided into negative control group(miR-NC),miR-421 mimics group,miR-421 mimics group+negative vector group(miR-421 mimics+vector)and miR-421 mimics group+overexpression plasmid group(miR-421 mimics+pcDNA-SFRP4),and transfected with negative control reagent,miR-421 analogs plus vectors and SFRP4 overexpression plasmid(pcDNA-SFRP4),respectively.The expression level of SFRP4 mRNA was detected by real-time quantitative PCR,and its effect on the proliferation of HEC-1A cells was detected by CCK-8 method.Results Microarray sequencing results showed that SFRP4 was one of differentially expressed genes and its expression was reduced in endometrial carcinoma(P<0.0001).SFRP4 expression affected the overall survival of patients with endometrial carcinoma(P<0.01),but was not related to the clinical stage(P>0.05).It was found that miR-421 may bind to SFRP4 using Targetscan software.Co

关 键 词：miR-421 SFRP4 子宫内膜癌 增殖 

分 类 号：R737.33[医药卫生—肿瘤]