Introduction: Our objective was to investigate the efficacy of all-trans retinoic acid (ATRA) alone and in combination chemotherapy with methotrexate (MTX) and combined with actinomycin D (Act-D) in choriocarcinoma cell culture models (JAR, JEG-3). Material and Methods: JAR and JEG-3 cells were cultured. ATRA, MTX and Act-D trial groups were determined with different doses. DMSO was applied as control group. Drugs were administered to the cells simultaneously, and 72 hours after drug administration, the cells were detached using trypsin-ethylenediamine tetraacetic acid solution. The degree of apoptosis was determined by flow cytometry. Statistical analyses of the apoptotic ratios were performed using SPSS 19.0 and the Wilcoxon test. Results: The ratio of apoptosis was statistically significant when only ATRA was applied on JAR and JEG-3 cell culture lines versus control group, p<0.05. The ratio of apoptosis was increased on JAR and JEG-3 cell culture lines, when ATRA was added in the combination of MTX 2 mu M, ACT-D 0.1 mu M, p<0.05. Discussion: ATRA increased the apoptotic ratios in both JAR and JEG-3 cell cultures. The apoptotic ratios were increased with the higher ATRA doses. The application of ATRA, MTX and Act-D combination on the JAR and JEG-3, cell line models, is made in both cell lines for the first time in the literature. The apoptotic data reveal that: ATRA could be used in choriocarcinoma treatment. Also, the combination of ATRA, MTX, and Act-D had stronger apoptotic effects than did the combination of MTX and Act-D. Therefore, ATRA also could be used as a synergistic drug and an option to combat the multi-drug resistance often encountered in the treatment of choriocarcinoma.