Oral treatment with the dual-receptor tyrosine kinase inhibitor AEE788 effectively reduces the number of peritumoral lymphatic vessels and the incidence of lymph node metastasis in nude mice with human HT29 colon cancer cells growing in the cecum. Whether inhibition of lymph node metastasis in colon cancer can be achieved by directly targeting lymphatic endothelial cells remains unclear. Using a microsurgical approach, we generated conditionally immortalized lymphatic endothelial cell lines from the H-2K(b)-tsA58 mouse mesentery and characterized these cells for the expression of lymphatic endothelial cell markers. Lymphatic endothelial cells were stimulated in culture with an array of tumor cell-produced cytokines, leading to the identification of redundant pathways for proliferation and survival. Treatment with AEE788 decreased the migration, proliferation, and survival of lymphatic endothelial cells, demonstrating that oral treatment with AEE788 effectively decreases the incidence of colon cancer lymphatic metastasis due, in part, to the direct inhibition of lymphatic endothelial cell signaling.