Abstract: The role of autophagy in the development and progression of hepatocellular carcinoma (HCC) is ambiguous and still little known. Autophagy stimulation may be of exceptional interest in antitumor pharmacotherapy of HCC. Rapamycin and lithium are typical inducers of autophagy. The aim of this study was to compare the level of autophagy in hepatocellular carcinoma-29 (HCC-29) cells after single and combined administration of lithium carbonate and rapamycin. Autolysosomes formation and significant increase of LC3 beta (+)– and LAMP1 (+)– autophagic structures were revealed in HCC-29 cells after lithium carbonate and rapamycin coadministration by transmission electron microscopy and immunofluorescence analysis. Using this combination of drugs may be a promising strategy for HCC chemotherapy, since it will allow the integration of various cellular signaling pathways that regulate autophagy and apoptosis in tumor cells.