EphB2调控claudin-6对子宫内膜癌侵袭和转移的影响  

EphB2 regulates the expression of claudin-6 to modulate endometrial canceRinvasion and metastasis

作　　者：孙帅 夏青 李凡[1,2] 闵锐 韩悦心 冯振中 李楠 SUN Shuai;XIA Qing;LI Fan;MIN Rui;HAN Yuexin;FENG Zhenzhong;LI Nan(Department of Pathology,Bengbu Medical College,Bengbu 233030,China;Department of Pathology,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233030,China;Department of Pathology,the Second Affiliated Hospital of Anhui Medical University,Hefei 230000,China;School of Clinical Medicine,Bengbu Medical College,Bengbu 233030,China)

机构地区：[1]蚌埠医学院病理学教研室,蚌埠233030 [2]蚌埠医学院第一附属医院病理科,蚌埠233030 [3]安徽医科大学第二附属医院病理科,合肥230000 [4]蚌埠医学院临床医学院,蚌埠233030

出　　处：《临床与实验病理学杂志》2023年第8期944-952,共9页Chinese Journal of Clinical and Experimental Pathology

基　　金：安徽高校自然科学研究项目(KJ2021A0701);蚌埠医学院研究生科研创新计划资助(Byycx21018);蚌埠医学院科技项目(2020byzd030)。

摘　　要：目的探讨EphB2和claudin-6的靶向关系,观察敲低和过表达EphB2基因通过调控claudin-6对子宫内膜癌细胞生长、迁移及侵袭的影响。方法采用TCGA数据库分析EphB2在子宫内膜癌和癌旁组织中的差异表达及其与临床病理特征的关系;应用GSEA法预测EphB2可能相关的功能通路;应用TCGA数据库分析EphB2互作基因;利用体外培养子宫内膜癌细胞;采用脂质体转染法转染子宫内膜癌细胞敲低和过表达EphB2基因;采用qRT-PCR法和Western blot法检测敲低和过表达EphB2各组细胞EphB2和claudin-6表达水平;提取转染后细胞进行细胞增殖、迁移和侵袭实验;免疫组化检测EphB2和claudin-6蛋白在子宫内膜样癌中的表达。结果TCGA数据库分析543例患者显示EphB2在子宫内膜癌中高表达,claudin-6可能是EphB2最具相关性基因(R=0.6);GSEA数据库分析显示EphB2表达可能与细胞黏附和细胞连接通路相关。qRT-PCR法显示干扰后EphB2 mRNA表达:阴性对照组:1.055±0.284、siRNA-EphB2组:0.499±0.264。划痕实验法检测不同分组中癌细胞的迁移能力,干扰后划痕愈合率显示:阴性对照组:0.629±0.02、空白对照组:0.676±0.006、siRNA-EphB2组:0.352±0.009;过表达后划痕愈合率显示:空白对照组:0.151±0.012、空载体组:0.271±0.015、EphB2过表达组:0.672±0.011;敲低EphB2后claudin-6表达显著下降,细胞增殖活力明显降低,细胞侵袭能力亦受到明显抑制,差异有统计学意义(P均<0.05);过表达EphB2后claudin-6表达明显上升,细胞迁移和侵袭能力亦明显增强,差异有统计学意义(P均<0.05)。实验显示:EphB2和claudin-6在子宫内膜样癌(128例)和癌旁组织(28例)中的表达,差异有统计学意义(P<0.05)。结论EphB2在子宫内膜癌中高表达,EphB2基因敲低和过表达影响子宫内膜癌细胞的生长及恶性生物学行为,其机制可能与调控claudin-6表达水平有关。Purpose To explore the targeting relationship between EphB2 and claudin-6,and to observe the effects of EphB2 gene knockdown and overexpression on the growth,migration,and invasion of endometrial canceRcells regulated by claudin-6.Methods The expression of EphB2 in endometrial canceRand adjacent tissues and its correlation with clinical pathological parameters were analyzed in TCGA database.The potential functional pathways of EphB2 in endometrial canceRwere predicted using the GSEA method.EphB2 interacting genes were identified through analysis of the TCGA database.Endometrial canceRcells were cultured in vitro,and EphB2 gene knockdown and overexpression were achieved by transfecting the cells using lipid-based transfection method.The expression levels of EphB2 and claudin-6 in each group of transfected cells were detected by qRT-PCRand Western blot.Transfected cells were used foRcell proliferation,migration,and invasion experiments.The protein expression of EphB2 and claudin-6 in endometrioid carcinoma was determined by immunohistochemistry.Results Analysis of data from 543 patients in the TCGA database revealed that EphB2 was highly expressed in endometrial cancer,and claudin-6 was likely the most correlated gene(R=0.6)to EphB2.GSEA database analysis indicates that EphB2 might be associated with cell adhesion and cell junction pathways.AfteRinterference,EphB2 mRNA expression was as follows:negative control group:1.055±0.284;siRNA-EphB2 group:0.499±0.264.Scratch assay was performed to assess the migration ability of canceRcells in different groups.The healing rates afteRinterference were as follows:negative control group:0.629±0.02;blank control group:0.676±0.006;siRNA-EphB2 group:0.352±0.009.The healing rates afteRoverexpression were:blank control group:0.151±0.012;empty vectoRgroup:0.271±0.015;EphB2 overexpression group:0.672±0.011.Knockdown of EphB2 significantly decreased claudin-6 expression,cell proliferation,and invasion capacity with statistical significance(P<0.05).Overexpression of EphB2 led t

关 键 词：子宫肿瘤 子宫内膜癌 EPHB2 claudin-6 生物学行为 

分 类 号：R737.33[医药卫生—肿瘤]