IFIT1通过激活Wnt/β-catenin通路促进胆管癌进展  

IFIT1 promotes progression of cholangiocarcinoma by activating Wnt/β-catenin pathway

作　　者：王鹏[1,2] 张迪 黄嘉槟[3] 刘均立 洪健 向国安 WANG Peng;ZHANG Di;HUANG Jiabin;LIU Junli;HONG Jian;XIANG Guoan(The Second School of Clinical Medicine,Southern Medical University,Guangzhou,510515,China;Department of Gas-trointestinal Surgery,Guangdong Second Provincial General Hospital,Guangzhou,510317,China;Department of Patho-physiology,School of Medicine,Jinan University,Guangzhou,510630,China.E-mail:[e-mail])

机构地区：[1]南方医科大学第二临床医学院,广东广州510515 [2]广东省第二人民医院胃肠外科,广东广州510317 [3]暨南大学基础医学与公共卫生学院,广东广州510632

出　　处：《中国病理生理杂志》2023年第5期769-778,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81871987);广东省基础与应用基础研究基金(No.2023A1515012905);广州市校(院)联合资助(登峰医院)基础研究项目(No.202201020304)。

摘　　要：目的:探讨干扰素诱导的三十四肽重复蛋白1(IFIT1)对胆管癌进展的影响及其调控机制。方法:RT-qPCR和Western blot法分别测定人正常胆管上皮细胞系HIBEpiC和6种不同分化程度胆管癌细胞系中IFIT1的mRNA和蛋白表达水平;检测人胆管癌和癌旁组织中IFIT1的表达水平,分析其与临床病理特征的相关性及预后价值。采用CCK-8、平板集落和Transwell实验检测IFIT1下调对胆管癌细胞增殖、迁移和侵袭的影响;选用胆管癌细胞系HuCCT1构建IFIT1稳定敲减的细胞株,分别采用裸鼠皮下移植瘤及肺转移瘤模型,观察IFIT1对胆管癌生长及转移的影响。通过公共数据库基因富集分析富集于IFIT1的相关信号通路,并进行验证。结果:相对于正常胆管上皮细胞和低侵袭性的胆管癌细胞系,高侵袭性的胆管癌细胞系中IFIT1呈高表达;人胆管癌组织中IFIT1表达水平显著高于癌旁组织(P<0.01),且与肿瘤恶性特征(肿瘤大小、淋巴结转移和肿瘤TNM分期)呈正相关,肿瘤组织中IFIT1高表达的患者总生存期相对较短(P<0.01)。IFIT1下调显著抑制胆管癌细胞的增殖、迁移和侵袭。在皮下移植瘤模型中,敲减IFIT1后皮下移植瘤生长速度减慢,体积缩小,重量减轻(P<0.01);在尾静脉肺转移瘤模型中,敲减IFIT1可显著减少裸鼠肺表面转移瘤结节数目(P<0.01)。生物信息学分析提示,与上皮-间充质转化(EMT)相关的Wnt/β-catenin通路在IFIT1高表达组富集。胆管癌细胞中敲减IFIT1可抑制Wnt/β-catenin通路,且EMT标志物vi⁃mentin和Snail表达减少。结论:IFIT1通过激活Wnt/β-catenin通路增强胆管癌细胞EMT,从而促进肿瘤进展。AIM:To explore the role of interferon-induced protein with tetratricopeptide repeats 1(IFIT1)in the development of cholangiocarcinoma(CCA).METHODS:RT-qPCR and Western blot were used to evaluate the ex⁃pression of IFIT1 at mRNA and protein levels,respectively,in various human cell lines,including a normal bile duct cell line and six distinct CCA cell lines.The level and prognostic value of IFIT1 were validated in human CCA and para-tumor tissues,accompanied by a comprehensive analysis of the pathological parameters and clinical significance of IFIT1.To as⁃certain the role of IFIT1 in CCA progression,we conducted CCK-8,colony formation and Transwell assays to investigate the impact of IFIT1 knockdown on the proliferation,migration and invasion of CCA cells in vitro.For in vivo experimenta⁃tion,we chose HuCCT1,a human CCA cell line with a high metastatic potential,to create subcutaneous xenograft and lung metastatic tumor models in nude mice.We observed the effect of IFIT1 on the growth and metastasis of orthotopic CCA xenograft with IFIT1 knockdown.Gene set enrichment analysis(GSEA)of public databases was performed to reveal the underlying mechanism regulated by IFIT1 in CCA progression,which was subsequently confirmed by functional studies in vitro.RESULTS:Compared with normal bile duct cells,IFIT1 was significantly up-regulated in human CCA cell lines,particularly in HuCCT1 and QBC939 cells with higher metastatic potential,as well as in CCA tissues compared with para-tumor tissues(P<0.01).Moreover,in a separate cohort of 44 CCA patients,up-regulated IFIT1 was positively corre⁃lated with tumor malignancy features such as tumor size,lymph node metastasis,TNM staging,and poor prognosis(P<0.01).These results suggest that IFIT1 plays a crucial role in CCA malignancy.Functional analysis revealed that knock⁃down of IFIT1 effectively inhibited the growth and metastasis of CCA cells in vitro and in vivo(P<0.01).Bioinformatic screening analysis indicated that the Wnt/β-catenin pathway,an epithelial-mesenchymal t

关 键 词：干扰素诱导的三十四肽重复蛋白1 胆管癌 上皮-间充质转化 WNT/Β-CATENIN信号通路 

分 类 号：R363.2[医药卫生—病理学] R735.8[医药卫生—基础医学]