miR-767-5p通过调控TBL1XR1蛋白表达抑制人卵巢癌细胞系侵袭  

miR-767-5p inhibits the invasion of ovarian cancer cell lines by regulating the expression of TBL1XR1 protein

作　　者：冷海娜[1] 张艳[1] 王晓栋 张学友 李洪利 LENG Haina;ZHANG Yan;WANG Xiaodong;ZHANG Xueyou;LI Hongli(Department of Gynaecology,Weifang Maternal and Child Health Care Hospital,Weifang 261000,China;College of Life Sciences and Technology,Weifang Medical College,Weifang 261000,China;School of Basic Medicine,Weifang Medical College,Weifang 261000,China)

机构地区：[1]潍坊市妇幼保健院妇科,山东潍坊261000 [2]潍坊医学院生命科学与技术学院,山东潍坊261000 [3]潍坊医学院基础医学院,山东潍坊261000

出　　处：《基础医学与临床》2023年第3期408-414,共7页Basic and Clinical Medicine

基　　金：山东省自然科学基金(ZR2019MH033)。

摘　　要：目的探索miR-767-5p和转导素β1X连锁受体蛋白1(TBL1XR1)对卵巢癌进展的影响以及作用机制。方法数据库分析:使用GEO、miRPATH数据库和RStudio对miR-767-5p进行KEGG(京都百科全书)富集分析,并在GEO数据集低表达和高表达microRNA中各选择差异表达倍数TOP5的microRNAs,按照差异倍数由低到高排列;miRDB等数据库预测miR-767-5p可结合的靶基因并取交集,结合Open Target Platform、GEPIA等数据库筛选出目的基因;GEPIA、THPA数据库分析TBL1XR1在人体各器官表达量分析、免疫组化分析以及生存曲线分析。实验验证:双荧光素酶报告基因实验验证miR-767-5p和TBL1XR1存在结合靶点;Western blot检测卵巢癌细胞系(OC3、SKOV-3、HO-8910)和正常卵巢上皮细胞系(IOSE80)中TBL1XR1的表达;在SKOV-3细胞中转入miR-767-5p过表达质粒与TBL1XR1过表达质粒后,Western blot检测TBL1XR1表达量,Tanswell小室法检测细胞侵袭。结果数据库分析:选择低表达差异倍数最高的miR-767-5p作为目的microRNA进行后续研究;miR-767-5p参与多种癌的相关通路,且miR-767-5p、TBL1XR1与乳腺癌、卵巢癌等联系密切;TBL1XR1在卵巢癌中呈现高表达,且与患者预后不良相关。实验验证:miR-767-5p和TBL1XR1可靶向结合(P<0.05);相对于IOSE80细胞,TBL1XR1在OC3、SKOV-3、HO-8910细胞中呈现明显高表达;转入miR-767-5p过表达质粒后,TBL1XR1表达量降低,卵巢癌细胞侵袭性降低(P<0.05),而同时转入TBL1XR1过表达质粒后可在一定程度上减弱这种影响。结论miR-767-5p通过调控TBL1XR1的表达抑制人卵巢癌细胞系的侵袭。Objective To explore the effect and mechanism of miR-767-5P and transductinβ1X linked receptor protein 1(TBL1XR1)on the progression of ovarian cancer.Methods Database analysis:KEGG(Kyoto Encyclopedia of Genes and Genomes,Kyoto Encyclopedia)enrichment analysis of miR-767-5P using GEO,miRPATHdatabase and RStudio TOP5 microRNAs with differential expression multiples were selected from low and high expression microRNAs in the GEO data set,then ranked from low to high in terms of differential multiples.The target genes for miR-767-5p were predicted by miRDB and other databases.The intersection was selected.The target genes were screened by the Open Target Platform,GEPIA and other databases.GEPIA and THPA databases were used to analyze the expression level of TBL1XR1 in human organs,immunohistochemical analysis and survival curve analysis.Experimental verification:Dual luciferase assay detected the binding target of miR-767-5P and TBL1XR1.Western blot was used to detect TBL1XR1 expression in ovarian cancer cell lines(OC3,SKOV-3,HO-8910)and normal ovarian epithelial cell line(IOSE80).After transferring into SKOV-3 cells with miR-767-5P over-expression plasmid and TBL1XR1 over-expression plasmid,Western blot was used to detect the expression level of TBL1XR1 and Tanswell cell assay was used to detect cell invasion.Results Database analysis:miR-767-5p with the highest differential ratio of low expression was selected as the target microRNA for follow-up research;miR-767-5p was involved in various cancer-related pathways,and miR-767-5p and TBL1XR1 were closely associated with breast cancer and ovarian cancer,etc.TBL1XR1 was highly expressed in ovarian cancer and was associated with poor prognosis.Experimental verification:miR-767-5p and TBL1XR1 could be targetly bound(P<0.05);Compared with IOSE80,TBL1XR1 was significantly over-expressed in OC3,SKOV-3 and HO-8910 cells.After the transfection of miR-767-5p over-expressed plasmid,the expression level of TBL1XR1 was decreased and the aggressiveness of ovarian cancer cells was

关 键 词：卵巢癌 侵袭 miR-767-5p 转导素β1X连锁受体蛋白1(TBL1XR1) 

分 类 号：R737.31[医药卫生—肿瘤]