Introduction: Tumor diseases are a leading cause of death worldwide and there is a rising need to discover efficient form of tumor treatment. Newly synthesized ferrocene derivatives show potential antitumor activity by inducing apoptosis in tumor cells without damaging healthy cells. Objective: The aim of this study was to examine the proapoptotic potential of ferrocene derivatives (Df-1, Df-2, Df-3, Df-4) on tumor cell lines and to compare the results of samples treated with ferrocene derivatives with a control, untreated sample. Materials and methods: Cells were treated with ferrocene derivatives for 72 h and the flow cytometry was used to detect the influence of ferrocene derivatives on cell lines in the process of apoptosis, and autophagy was determined. The obtained data are presented as a percentage of the population, using a statistical function for data analysis and results processing obtained from flow cytometry in the FlowJо program. Results: The results indicate a significant statistical effect of ferrocene derivatives Df-3 and Df-4 on CaCo-2, CCRF-CEM and Raji cell lines in the process of apoptosis and late apoptosis/necrosis. A significant statistical effect of ferrocene derivatives Df-3 on induced autophagy in CCRF-CEM cell line, while Df-1, Df-2, and Df-3 derivatives showed an inhibitory effect on autophagy in the CaCo-2 cell line. Conclusion: Based on the obtained results, it can be concluded that the derivatives Df-3 and Df-4 show a proapoptotic effect in apoptosis and late apoptosis/necrosis. The Df-3 derivative showed the highest proapoptotic effect of all tested derivatives. Research of the influence of ferrocene derivatives in the process of autophagy needs to be further investigated, using other methods.