Hepatic tumors result in impaired ammonia metabolism, leading to a tumor microenvironment (TME) characterized by elevated ammonia levels. While normal cells are inhibited in this high-ammonia environment, the specific mechanisms underlying ammonia cytotoxicity remain unclear. In this study, we employed embryonic cells and isolated primary hepatocytes, utilizing CCK-8 assays, Western blotting, and qPCR techniques to investigate the toxic effects of ammonia. Our findings indicate that elevated ammonia levels significantly inhibit cell survival and mTORC2 signaling, while simultaneously inducing apoptosis and endoplasmic reticulum stress. These findings enhance the understanding of the physiological functions of normal cells within TMEs.