摘要
目的探究成纤维细胞活化蛋白(fibroblast activation protein,FAP)在食管鳞癌(esophageal squamous cell carcinoma,ESCC)中的表达水平及对其预后的影响,进一步研究肿瘤相关成纤维细胞(cancer-associated fibroblasts,CAFs)中FAP的表达对ESCC恶性进展的影响。方法首先,通过TCGA数据库评估FAP在31种肿瘤的癌和癌旁组织中表达水平的差异,并分析其对ESCC患者预后的影响;进一步收集高发区ESCC患者癌和癌旁新鲜组织用于原代CAFs和癌旁正常成纤维细胞(normal fibroblasts,NFs)的提取和分离,利用qRT-PCR、Western blot和细胞免疫荧光进行表型鉴定和功能分析;收集NFs、CAFs和经FAP抑制剂(FAPi)处理后的CAFs细胞上清,与KYSE150细胞构建体外共培养模型,通过EdU法、CCK-8法、平板克隆、Transwell和划痕实验探究FAP表达水平对KYSE150细胞增殖、侵袭和迁移等恶性行为的影响;最后,通过构建食管癌小鼠皮下荷瘤模型,在体内探究CAFs中FAP的表达对ESCC恶性进展的影响。结果在包含ESCC在内的10种恶性肿瘤中,癌组织中FAP的表达均显著高于癌旁组织,且ESCC中FAP的高表达提示患者的不良预后,并且FAP可作为ESCC患者预后的独立危险因素;从ESCC及其癌旁组织中成功提取、分离出原代CAFs和NFs,经鉴定表型正确;体外共培养结果显示,与NFs相比,CAFs高表达FAP并显著促进ESCC细胞的增殖、侵袭和迁移能力,而FAPi处理可显著抑制上述恶性行为;体内实验证实FAPi和CAFs去激活处理,对荷瘤小鼠肿瘤生长具有显著抑制作用。结论在ESCC中,FAP的高表达与CAFs的活化和患者的不良预后呈正相关,促进肿瘤的恶性进展。
Objective To investigate the expression level of fibroblast activation protein(FAP)in esophageal squamous cell carcinoma(ESCC)and its prognostic implications,and to further explore the role of FAP expression in cancer-associated fibroblasts(CAFs)in driving the malignant progression of ESCC.Methods The expression levels of FAP in tumor and adjacent tissues of 31 types of tumors were assessed by utilizing the TCGA database,followed by an analysis of its influence on the prognosis of ESCC.Fresh tumor and normal tissue samples were acquired from ESCC patients in high-incidence regions for extracting and purifying primary CAFs and normal fibroblasts(NFs).Phenotypic identification and functional analyses were conducted via qRT-PCR,Western blot,and immunofluorescence.Supernatants from NFs,CAFs,and CAFs treated with the FAP inhibitor(FAPi)were collected to establish an in vitro co-culture model with KYSE150 cells.The impact of differential FAP expression levels on the malignant progression of KYSE150 cells,encompassing proliferation,invasion,and migration were assessed by using EdU assay,CCK-8 assay,colony formation assay,Transwell assay,and scratch assay.The in vivo expression of FAP in CAFs was detected in a subcutaneous tumor model of esophageal cancer in mice to elucidate its role in the malignant progression of ESCC.Results In 10 types of malignant tumors,including ESCC,the expression of FAP was markedly elevated in tumor tissues compared to adjacent non-tumorous tissues.Moreover,elevated FAP expression in ESCC was associated with a poor prognosis and may serve as an independent prognostic risk factor for ESCC patients.Primary CAFs and NFs were effectively isolated from ESCC tumor and adjacent tissues,and their phenotypic characteristics were precisely characterized.The results of co-culture experiments indicated that,in comparison to NFs and untreated cells,CAFs with elevated FAP expression markedly enhanced the proliferation,invasion,and migration of KYSE150 cells.Conversely,FAPi treatment significantly suppressed these malignant behaviors.In vivo experiments had demonstrated that the deactivation treatment of FAPi and CAFs exerted a significant inhibitory effect on tumor growth in tumor-bearing mice.Conclusion In ESCC,the high expression of FAP is positively correlated with the activation of CAFs and the poor prognosis of patients,thereby promoting the malignant progression of the tumor.
作者
李佳怡
张理
刘书培
陈攀
张小漫
牛浩瑾
杨平娟
高震东
石林林
高社干
Li Jiayi;Zhang Li;Liu Shupei(Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment,Henan Key Laboratory of Cancer Epigenetics,the First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471003,China;Department of Clinical Laboratory,the First Af filiated Hospital of Henan University of Science and Technology,Luoyang 471003,China)
出处
《华中科技大学学报(医学版)》
北大核心
2025年第1期13-21,共9页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.82302966)
河南省医学科技攻关省部共建重点项目(No.SBGJ202102199,No.LHGJ20230455)
河南省省级科技研发计划联合基金资助项目(No.232103810048)
洛阳市社会发展公益专项(No.2302016A,No.2302016Y)。
关键词
食管鳞癌
成纤维细胞活化蛋白
肿瘤相关成纤维细胞
esophageal squamous cell carcinoma
fibroblast activation protein
cancer-associated fibroblast
