Heliomycin is a natural compound extracted from the metabolites of Actinomycetes bacteria. We aim to evaluate the anticancer role of Heliomycin, against Tamoxifen as a standard drug. Flow cytometry and analysis of the cell cycle to detect the ratios of apoptosis, and cell cycle arrest were performed, and the genes expression was assayed using real-time RT-PCR. CK-19 protein was assessed using immunocytochemistry. Also, cytopathological alterations were evaluated using H&E stain and electron microscope examination. qRT- PCR showed that HER-2 expression was less elevated in tamoxifen-treated cells than in heliomycin-treated cells in the control cell. For P53 it showed significant up-regulation in heliomycin-treated cells than tamoxifen-treated cells compared to the control. In contrast, ERα, TNFα, and TLR-4 showed the reverse. The flow cytometry assay revealed a considerable increase in early and late apoptosis in the treated group with Tamox. and Helio. compared with the control. H&E stain and ultrastructure study showed cell apoptosis and chromatin degradation and cell shrinkage in treated cells against control. Immunohistochemically, CK-19 showed a severe reaction in control cells while a mild reaction in heliomycin-treated cells versus a moderate reaction in tamoxifen-treated cells. Heliomycin possesses accepted anticancer behaviour against breast cancer MCF-7 cells, as it activates cancerous cell apoptosis with upregulation of tumour suppressor genes.