In this paper, the results showed that the proliferation activity of T24 cells and 5637 cells treated with bupivacaine (0.25–16 mM) was significantly inhibited, and our study also confirmed that bupivacaine promoted apoptosis and ferroptosis in bladder cancer cells by inhibiting PI3K/AKT/mTOR signaling pathway. The gene discussed is MTOR; the disease is urinary bladder carcinoma.