ObjectiveCholangiocarcinoma, a cancer of the biliary duct, is frequently diagnosed in advanced stages with poor prognosis. Understanding the molecular mechanisms driving its progression is crucial for developing effective treatment approaches. This research delves into the role of LINC00313 in regulating the aggressiveness of cholangiocarcinoma cells.MethodsClinical samples were collected to examine the expression pattern of LINC00313, miR-320b, and MITF using RT-qPCR and western blot analysis. Functional experiments, including cell proliferation, migration, and invasion assays, as well as apoptosis detection, were performed to assess the malignant properties of cholangiocarcinoma cells. The tumorigenic potential of cholangiocarcinoma cells was further investigated in a xenograft mouse model.ResultsElevated levels of LINC00313 were detected in both cholangiocarcinoma tumors and cell lines. Knocking down LINC00313 reduced the aggressiveness of cholangiocarcinoma cells and hindered their ability to form tumors in vivo. It was discovered that miR-320b is a target of LINC00313, exhibiting an expression pattern and functional role that contrasts with LINC00313. Moreover, the LINC00313/miR-320b axis was found to regulate the expression of MITF, thereby influencing the aggressiveness of cholangiocarcinoma cells.ConclusionLINC00313/miR-320b/MITF axis is implicated in the malignant progression of cholangiocarcinoma, indicating the potential of targeting LINC00313 as a strategy for cholangiocarcinoma clinical management.