The Role And Mechanism Of Forkhead Box Protein M1 In Intrahepatic Cholangiocarcinoma

Objective(s):Intrahepatic cholangiocarcinoma(ICC)is highly invasive and has a poor prognosis.Its pathogenesis involves many aspects,such as genetic mutations and dysregulation of cell signaling pathways.The purpose of this study was to explore the expression of FoxM1 in ICC and its influence on the biological behavior of ICC cells,and to analyze the relevant signaling pathways affected by the abnormal expression of FoxM1 in ICC through bioinformatics,so as to provide theoretical basis for clinical diagnosis and treatment of ICC.Method(s):1.The Cancer Genome Atlas(TCGA)data were collected,and the expression of FoxM1 in ICC tissues and normal control tissues was compared at the m RNA level,and the relationship between the expression of FoxM1 and the tumor mutation load score and prognosis of ICC patients was analyzed;2.Western blotting was used to detect the expression level of FoxM1 protein in ICC cell lines,and lentivirus transfection was used to establish stable up-regulated and down-regulated expression of FoxM1 in ICC cell lines;MTT test and Transwell cell invasion test were used to detect its effect on the proliferation and invasion of ICC cells;The expression levels of MMP-9 and MMP-2 m RNA in stably transfected ICC cell lines were detected by q PCR.3.Collect data from the public database(TCGA and KEGG),divide them into FoxM1 high expression group and low expression group according to the median of the overall FoxM1 m RNA expression value,and compare the relevant signal pathways that may be affected between different expression groups of FoxM1 through Gene Set Enrichment Analysis(GSEA);At the same time,the relationship between the expression of FoxM1 and the level of P53 m RNA in ICC cell lines was compared by q PCR.Result(s): 1.Bioinformatics analysis showed that the expression of FoxM1 in ICC tumors was significantly higher than that in normal tissues(P<0.0001).The expression of FoxM1 was related to the tumor mutation burden score of ICC(P=0.021),but not to the survival and prognosis of ICC(P=0.308);2.The expression level of FoxM1 protein in SSP-25 cell line was the highest,while that in HCCC-9810 cell line was the lowest(P<0.01 for comparison between the two groups,P<0.0001 for analysis of variance);After lentivirus transfected FoxM1 to construct stable transfected ICC cell line,overexpression of FoxM1 can enhance the proliferation and invasion ability of ICC cells,while down-regulation of FoxM1 expression can inhibit the growth and invasion ability of ICC cells.Overexpression of FoxM1 may enhance the expression of MMP-9 and MMP-2 m RNA in ICC cells(all P < 0.05);3.GSEA analysis showed that when FoxM1 was highly expressed in ICC,there were significant differences in the expression of many gene signal pathways,such as P53 and PLK1(FDR-q values < 0.250).q PCR results showed that the expression of P53 m RNA in HCCC-9810-FoxM1 cell group with up-regulated FoxM1 expression after lentivirus transfection was significantly higher than that in the control group,while the expression of P53 m RNA in SSP-25-sh FoxM1 cell group with down-regulated FoxM1 expression was lower than that in the control group(P < 0.0001).Conclusion(s):1.FoxM1 promotes the proliferation and invasion of ICC cells.2.FoxM1 may be involved in regulating the expression of MMP-9 and MMP-2 in ICC cells.3.The expression of FoxM1 in ICC may be related to P53 signal pathway.