Abstract: The current study was to investigate the joint toxicity of zearalenone(ZEA)and aflatoxin B1(AFB₁)on HepG2 cells. Changes in multi-parameter cell biology indicators and expression of apoptosis-related target genes were performed,and the combined toxicity model was used to evaluate the combined toxicity pattern. The results demonstrated that,ZEA and AFB₁ had dose- and time-dependently decreased with the viability of HepG2 cells,and the IC₅₀ values was 78.57 and 18.04 μmol/L at 24 h respectively. Compared with the individual group,the combined group could significantly(P<0.05)reduced cell viability and increased cell death rate,membrane permeability,intracellular calcium level,reactive oxygen species(ROS),and DNA damage. The results of qRT-PCR showed that,ZEA and AFB₁ could induce apoptosis in HepG2 cells by significantly(P<0.05)increasing the mRNA expression levels of p53,Caspase-3 and Bax,and significantly(P<0.05)decreasing the expression of Bcl-2.The combined toxicity modes of ZEA and AFB₁ were evaluated according to the additive effect model,which showed synergistic effect.