NLRC3对子宫内膜样癌作用的研究  

Study on the effect of NLRC3 on endometrioid carcinoma

作　　者：孙一卿 申晓畅 颜磊 赵兴波[1] Sun Yiqing;Shen Xiaochang;Yan Lei(Department of Obstetrics and Gynecology,Provincial Hospital Affiliated to Shandong University,Jinan 250021;Center for Reproductive Medicine,Reproductive Hospital Affiliated to Shandong University,Jinan 250001)

机构地区：[1]山东大学附属省立医院妇产科,济南250021 [2]山东大学附属生殖医院生殖医学中心,济南250001

出　　处：《现代妇产科进展》2022年第11期820-824,共5页Progress in Obstetrics and Gynecology

基　　金：山东省自然科学基金项目(No:ZR2018MH207)。

摘　　要：目的:探讨NLR家族含CARD结构蛋白3(NLRC3)在子宫内膜癌组织中的表达情况及其对子宫内膜癌细胞系生物学行为的影响。方法:采用GEPIA数据库分析子宫内膜癌患者NLRC3表达水平及其与无病生存率的关系。免疫组化、Western blot法检测NLRC3在子宫内膜样癌组织、癌旁组织、对照组子宫内膜组织中的表达。Ishikawa(ISK)、KLE细胞转染过表达NLRC3慢病毒,细胞平板克隆、划痕愈伤实验、Transwell迁移及侵袭实验检测NLRC3对体外细胞增殖、迁移、侵袭的影响。Western blot实验检测过表达NLRC3对上皮间质转化(EMT)的影响。构建裸鼠皮下成瘤模型,检测过表达NLRC3对肿瘤生长的影响。结果:GEPIA数据库分析显示,NLRC3在子宫内膜癌组织中低表达,较高水平NLRC3与较好的无病生存率有关(P<0.05)。免疫组化、Western blot实验结果与数据库分析一致,NLRC3在子宫内膜样癌组织中的表达水平低于正常子宫内膜组织、配对癌旁组织(P<0.05),NLRC3表达随肿瘤病理分化程度降低而逐渐降低。ISK、KLE细胞过表达NLRC3后,克隆形成率下降(P均<0.05),细胞划痕愈合率下降(P_(ISK)<0.01,P_(KLE)<0.0001),细胞穿膜数减少(P均<0.01)。Western blot检测EMT相关指标(E-cadherin、N-cadherin、Vimentin),KLE细胞中N-cadherin、Vimentin表达水平下降(P<0.05)。裸鼠皮下成瘤模型表明,肿瘤重量降低(P<0.05)。结论:NLRC3在子宫内膜样癌组织中低表达,可抑制子宫内膜癌细胞系的增殖、迁移、侵袭、EMT等恶性行为。Objective:To investigate the expression of NLR family CARD-containing structural protein 3(NLRC3)in endometrial cancer tissues and its effect on the biological behavior of endometrial cancer cell lines.Methods:The GEPIA database was used to analyze the expression level of NLRC3 in patients with endometrial cancer and its relationship with disease-free survival.The expression of NLRC3 in endometrioid carcinoma tissues,adjacent tissues and control endometrial tissues was detected using Immunohistochemistry and Western blot.Ishikawa(ISK)and KLE cells were transfected with overexpressing NLRC3 lentivirus,and the effect of NLRC3 on cell proliferation,migration and invasion in vitro was detected by cell plate cloning,scratch callus assay,Transwell migration and invasion assay.Western blot assay was used to detect the effect of NLRC3 overexpression on epithelial-mesenchymal transition(EMT).A subcutaneous tumorigenesis model in nude mice was constructed to detect the effect of overexpression of NLRC3 on tumor growth in vivo.Results:The GEPIA database analysis showed that NLRC3 was lowly expressed in endometrial cancer tissues,and higher levels of NLRC3 were associated with better disease-free survival(P<0.05).The results of immunohistochemistry and Western blot experiments were consistent with the analysis of the database.The expression in endometrioid carcinoma tissue was lower than that in the control endometrium and the paired adjacent tissue(P<0.05),the expression of NLRC3 gradually decreased with the decrease of tumor pathological differentiation.After ISK and KLE cells overexpress NLRC3,the colony formation rate decreased(P<0.05),the wound healing rate of cells decreased(P_(ISK)<0.01,P_(KLE)<0.0001),the number of cells penetrating membrane decreased(both P<0.01).EMT related indicators(E-cadherin,N-cadherin,Vimentin)were detected by Western blot,and the expression levels of N-cadherin and Vimentin in KLE cells decreased(P<0.05).Subcutaneous tumorigenesis model in nude mice showed that tumor weight decreased(P<0.05).

关 键 词：子宫内膜样癌 NLRC3 迁移 侵袭 EMT 

分 类 号：R737.33[医药卫生—肿瘤]