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. Here, MTOR is linked to urinary bladder cancer.