In summary, our results demonstrated that upregulation of LINC00665 was involved in gemcitabine-resistance of CCA cells, and this was due to ceRNA regulation of miR-424-5p/BCL9L axis, thus activated Wnt/β-Catenin signaling and its downstream EMT and stemness related genes, and eventually promoted EMT and stemness properties of CCA cells. The gene discussed is LINC00665; the disease is cholangiocarcinoma.