Proteasomes are essential for numerous cellular processes, including the cell cycle, regulation of gene expression and responses to cellular stress. Proteasome inhibitors are promising anticancer agents. The proteasome inhibitor bortezomib effectively suppresses certain types of cancer, including multiple myeloma and mantle cell lymphoma. However, bortezomib poorly inhibits solid tumors, including hepatocellular carcinoma. The activation of Akt represents an adaptive response that prevents bortezomib-induced cell apoptosis. In the present study, bortezomib induced phosphorylation of EGFR, Src and Akt in hepatoma cells and inhibition of Src reduced bortezomib-induced EGFR and Akt phosphorylation. Treatment of hepatoma cells with bortezomib led to an increase in the levels of intracellular reactive oxygen species (ROS). The ROS scavenger N-acetyl-L-cysteine inhibits bortezomib-induced ROS production and abrogates the phosphorylation of Src, epidermal growth factor receptor and Akt. The combination of bortezomib and saracatinib, a Src inhibitor, synergistically induced hepatoma cell apoptosis. The present study concluded that ROS mediated the activation of the Src-EGFR-Akt cascade by bortezomib. The combination of the Src inhibitor and bortezomib holds promise in the treatment of hepatocellular carcinoma.