Eleutheroside E (EE) is a monomer compound isolated from Acanthopanax senticosus, which has functions of antifatigue, antioxidation, and immune regulation. However, the function of EE in inhibiting cancer has rarely been reported. In this study, we used bioinformatics to determine the role and mechanisms of EE in inhibiting breast cancer (BRCA), prostate cancer (PRAD), lung cancer (LUAD), and bladder cancer (BLCA). Finally, the MTT method was used to evaluate the inhibitory effect of EE on breast cancer MDA-MB-231 cells, and the target genes were determined by qRT-PCR. The results showed that the survival rate of MDA-MB-231 cells was reduced by 27.33% after 400 μg/mL EE treatment. The results of qRT-PCR showed that EE down-regulated target genes in breast cancer cells, which was consistent with our predicted results. Moreover, our results indicated that EE may have impact on the transcription and translation of BLCA cells by targeting ADCY2, ADCY5 and JUN. In BRCA, we identified three targets for EE. In addition, EE affected DNA repair and the development of BRCA by affecting the expression of RAD51. In LUAD, we identified 9 targets for EE, 8 of which were related to DNA repair. EE inhibited PRAD by targeting ADCY8. Our research was the first report on the targets and mechanisms of EE for suppressing cancer combined with clinical data. Our research also provides theoretical support for using EE to suppress cancer.