In this study, our results showed that Mus81 knockdown by lentivirus‐mediated shRNA could remarkably improve the sensitivity of common chemotherapeutic drugs including EPI, 5‐FU, MMC, and cisplatin in human HCC HepG2 and Bel‐7402 cells, and the RI value of EPI was the highest among the four agents we tested. Here, MUS81 is linked to hepatocellular carcinoma.