Zearalenone (ZEA) has long been recognized as a xenoestrogen, while the endocrine disrupting effects of aflatoxin B1 (AFB1) have been identified recently. Due to co-occurrence and endocrine disrupting potentials of ZEA and AFB1, it was hypothesized that co-exposure to ZEA and AFB1 might affect breast cancer cell growth. Consequently, the aim of this study was to evaluate the combined effects of ZEA and AFB1 (1 nM–100 nM) on cell growth and cell cycle progression, using a human breast cancer cell line MCF-7. Our results showed that ZEA and AFB1 produced significant interactive effects on cell growth, DNA synthesis and cell cycle progression. While ZEA promoted growth, DNA synthesis and cell cycle progression, AFB1 was cytotoxic and counteracted the effects of ZEA. ZEA altered the expression of several breast cancer related genes, whereas AFB1 had minimal effects on gene expression. With the use of specific inhibitors, ERα, GPER and MAPK pathways were found to be responsible for ZEA’s effects on cell growth; while MAPK pathways might be involved in cytotoxic effects by AFB1. This study is first to report the effects of co-exposure of ZEA and AFB1 on breast cancer cell growth, possibly through ER dependent pathway. This suggested that endocrine-disrupting mycotoxins that co-occur in human food can interact and influence human health. Future work on interactive effects of endocrine-disrupting mycotoxins or other xenoestrogens is warranted, which will contribute to improved risk assessments.