IFIT1 Promotes The Progress Of Cholangiocarcinoma By Activatingwnt/β-catenin Pathway

Purpose and significance:The frequency of cholangiocarcinoma(CCA),the second most frequent malignant liver tumor,has gradually increased worldwide.Most patients are diagnosed as advanced,and the treatment effect at this time is not optimistic.Interferon-induced protein with tetrapeptide repeats 1(IFIT 1)has been reported to play a pivotal role in tumor progression in many cancer types.However,the role of IFIT1 in solid tumors,especially cholangiocarcinoma(CCA),remains unexplored.Here,we aim to investigate the function and regulatory mechanism of IFIT1 in CCA progression,and provide a theoretical basis for developing early screening markers and therapeutic targets of CCA.Methods:Through bioinformatics analysis,the difference between the expression of IFIT1 in the important inducing factors of CCA and normal bile duct tissues was analyzed.Two models of spontaneous CCA induction in mice were constructed,and the expression of IFIT1 in different stages of CCA development was explored by immunohistochemistry(IHC).The tissues of 163 patients with CCA were collected to explore the expression level of IFIT1 in human CCA and adjacent tissues was detected by Western Blotting and IHC,and its correlation with clinicopathological features and prognostic value were analyzed CCK-8,colony formation,cell apoptosis,Wound-Healing and Transwell assays were used to detect the effects of down-regulation or up-regulation of IFITl expression on the proliferation,migration and invasion of CCA cells.To construct a stable HuCCTl cell line with IFIT1 knocked out,and apply it to nude mice subcutaneous tumor model to detect the effect of IFIT1 on the growth of CCA cells.Genetic enrichment analysis(GSEA)in public database used to obtain IFIT1-related signal pathways,and verify them.Results:Bioinformatics analysis showed that the expression of IFIT1 was higher in hepatitis B and nonalcoholic fatty liver than in normal bile duct tissues.At the same time,in the different stages of CCA carcinogenesis,it was found that the expression of IFIT1 showed a dynamic increase in both spontaneous rat and mouse models.By evaluating the tissues of 163 patients with CCA we found that the expression of IFIT1 in human CCA tissue was higher than that in normal bile duct tissue,and the expression of IFIT1 was related to lymph node metastasis and TNM staging of tumor,and the high expression of IFIT1 in tumor further predicted the short overall survival of CCA patients(P<0.01).In vitro cell experiments show that knocking out IFIT1 can effectively reduce the proliferation,migration and invasion of CCA cells,while overexpressing EFIT1 can promote the proliferation,migration and invasion of CCA cells.In vivo nude mice experiments showed that knocking out IFIT1 inhibited the subcutaneous tumorigenesis of CCA cells in nude mice.In mechanism,IFIT1 promotes epithelial-to-mesenchymal transition(EMT)of CCA cells by activating Wnt/β-catenin signaling pathway,which leads to increased invasiveness of CCA.This process has been confirmed in human CCA tissues.Conclusion:IFIT1 can promote EMT of CCA by activating intracellular Wnt/β-catenin pathway,and then regulate the progress of CCA.