Hepatocellular carcinoma (HCC) is one of the most commonly malignant tumors worldwide, characterized by the presence of many heterogeneous molecular cell events that contribute to tumor growth and progression. Endocytic processes are intimately involved in various pathological conditions, including cancer, since they interface with various cellular signaling programs. The ability of lithium to induce cell death and autophagy and affect cell proliferation and intracellular signaling has been shown in various experimental tumor models. The aim of this study was to evaluate the effects of lithium on vesicular transport in hepatocellular carcinoma cells. Using transmission electron microscopy we have characterized the endocytic apparatus in hepatocellular carcinoma-29 (HCC-29) cells in vivo and detailed changes in endocytotic vesicles after 20 mM lithium carbonate administration. Immunofluorescent analysis was used to quantify cells positive for EEA1-positive early endosomes, Rab11-positive recycling endosomes and Rab7-positive late endosomes. Lithium treatment caused an increase in EEA1- and Rab11-positive structures and a decrease in Rab7-positive vesicles. Thus, lithium affects diverse endocytic pathways in HCC-29 cells which may modulate growth and development of hepatocellular carcinoma.