摘要
目的:研究EMR2在肝外胆管细胞癌(extrahepatic cholangiocarcinoma,EHCC)中的表达及临床意义、生物学功能及机制。方法:采用免疫组化(immunohistochemistry,IHC)实验检测EMR2在EHCC组织及癌旁组织中的表达情况,并分析EMR2表达与EHCC患者临床病理特征之间的关系。利用Kaplan-Meier方法、COX比例风险模型分析EMR2表达在EHCC患者预后中的作用以及对EHCC患者预后造成危险的因素。通过qRT-PCR和Western blot实验测定EMR2在EHCC细胞系(QBC939、SK-CHA-1、KMBC)和正常胆管上皮细胞HIBEpic中mRNA及蛋白的相对表达水平,检验细胞敲低和过表达的效率。Western blot实验检测EMR2在上皮间质转化(epithelial-mesenchymal transformation,EMT)中的表达变化。细胞侵袭、增殖、迁移能力分别用Transwell侵袭实验、CCK-8实验、细胞划痕及Tranwell迁移实验检测。结果:IHC结果显示,EMR2在癌旁组织中阳性表达率低于EHCC组织,EMR2的表达与EHCC分化程度有关。Kaplan-Meier生存曲线显示:EMR2的表达与EHCC患者的预后有相关性,EMR2表达和EHCC分化程度是影响EHCC患者预后的独立危险因素。通过qRT-PCR和Western blot实验确定了以EMR2高表达细胞株QBC939和低表达细胞株KMBC作为后续实验对象,并且验证了转染效率。EMR2在EHCC细胞中的表达与细胞增殖、迁移、侵袭能力和N-cadherin、Vimentin蛋白水平成正比,与E-cadherin蛋白水平成反比。结论:EMR2表达与EHCC分化程度、预后有相关性,EMR2表达和分化程度是独立影响EHCC预后的因素。EMR2高表达能够促使EHCC细胞的增殖、侵袭、迁移能力增加,其机制可能与EMT有关。
Objective:To study the expression and clinical significance,biological function and mechanism of EMR2 in extrahepatic cholangiocarcinoma(EHCC).Methods:Immunohistochemistry(IHC)assay was used to detect the expression of EMR2 in EHCC tissues and paracancerous tissues and the relationship between EMR2 expression and the clinicopathological characteristics of EHCC patients was analyzed.Kaplan-Meier method and COX proportional hazard model were used to analyze the role of EMR2 expression in prognosis of EHCC patients and the risk factors for prognosis of EHCC patients.The relative expression levels of EMR2 mRNA and protein in EHCC cell lines(QBC939,SK-CHA-1,KMBC)and normal bile duct epithelial cells HIBEpic were determined by qRT-PCR and Western blot assays.The efficiency of cellular knockdown and overexpression was examined.Western blot assay detected the expression changes of EMR2 in epithelial-mesenchymal transformation(EMT).Cell invasion,proliferation and migration ability were examined by Transwell invasion assay,CCK-8 assay,cell scratch and Transwell migration assay,respectively.Results:IHC results showed that the positive expression rate of EMR2 in paracancerous tissues was lower than that in EHCC tissues,and the expression of EMR2 was related to the degree of differentiation of EHCC.Kaplan-Meier survival curves showed that the expression of EMR2 correlated with the prognosis of EHCC patients,and the expression of EMR2 and the degree of differentiation of EHCC patients were independent risk factors affecting the prognosis of EHCC patients.qRT-PCR and Western blot experiments confirmed that the high expression cells QBC939 and low expression cells KMBC of EMR2 were taken as the follow-up experimental objects,and the transfection efficiency was verified.The expression of EMR2 in EHCC cells was positively correlated with the ability of cells proliferation,migration,invasion and the protein levels of N-cadherin and Vimentin,and inversely correlated with E-cadherin protein level.Conclusion:EMR2 expression was correl
作者
苏莉莉
周帅
梁晚晴
周雪丽
韩啸
SU Lili;ZHOU Shuai;LIANG Wanqing;ZHOU Xueli;HAN Xiao(Department of Oncology,the First Affiliated Hospital of Bengbu Medical College,Anhui Bengbu 233000,China;Department of Hepatobiliary Surgery,the First Affiliated Hospital of Bengbu Medical College,Anhui Bengbu 233000,China;Department of Surgical Oncology,the First Affiliated Hospital of Bengbu Medical College,Anhui Bengbu 233000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第2期299-304,共6页
Journal of Modern Oncology
基金
安徽省自然科学基金项目(编号:1608085MH201)。
关键词
EMR2
肝外胆管细胞癌
免疫组化
EMR2
extrahepatic cholangiocarcinoma
immunohistochemistry
