Objective: To analyze the biological functions and its clinical significance of let-7b-5p target gene in cholangiocarcinoma utilizing bioinformatics. Methods: The paper focuses on the let-7b-5p target gene, and predicts its biological functions as well as related signal pathways through GO biological function and KEGG signal pathway enrichment analysis. The STRING database and Cytoscape are used to construct a protein-protein interaction network to screen core genes. Results: The results of GO analysis showed that let-7b-5p target gene was mainly enriched in biological processes such as Small GTPase binding, Rho GTPase binding, and Rac GTPase binding. The results of KEGG analysis showed that let-7b-5p target gene was significantly enriched in key signaling pathways including Focal adhesion and ECM-receptor interaction. Through protein-protein interaction network and module analysis, CXCL8 and SDC2 were screened as the core site. Conclusion: let-7b-5p can participate in the regulation of biological functions of tumor cells in cholangiocarcinoma, suggesting that it may play an important role as a tumor suppressor gene and biomarker in the occurrence and development of cholangiocarcinoma, which provides new ideas for its diagnosis and treatment.