Background: Piper retrofractum (PR) is traditional medicine in the South-East country. Objectives: This study examined the PR effects on anticancer and anti-migratory activity in human breast cancer cells. Materials and Methods: Determination of cancer cell death by the sulforhodamine B, colony formation, flow cytometry methods. Mechanism-induced cells death was assessed by intracellular reactive oxygen species generation, caspase 3 activity, mitochondrial function, gene expression, and protein expression. Results: The ethanolic extract of PR had higher effects against MCF-7 cells than DW extract expressing the high levels of piperine. The extracts stimulated cell death in MCF-7 cells and induced apoptosis with increasing caspase 3 activity, ROS generation, and decreasing mitochondria function. Moreover, the extract also inhibited gene-related cell proliferation, cdk2, cdk4, and ckd6, and altered protein-related cell growth, cyclin D1 reduction and p21 induction. Finally, the extract caused inhibition of cells migration by reducing MMP 9 levels. The extracts of PR stimulated MCF-7 cells death in association with increasing apoptosis and inhibiting cancer cells migration. The induction of cancer cell death may be through modulating mitochondrial function. Conclusion: These extracts may be a novel strategy to improve the efficacy of chemotherapy to treat MCF-7.