Bupivacaine modulates the apoptosis and ferroptosis in bladder cancer via phosphatidylinositol 3-kinase (PI3K)/AKT pathway

ABSTRACT

The study aimed to explore the effects of local anesthetic bupivacaine on bladder cancer cells in vivo and in vitro. The cytotoxicity was detected by MTT assay. Apoptosis was measured by Hoechst 33342 staining and TUNEL. The contents of Fe²⁺, Malondialdehyde (MDA), Glutathione (GSH) and reactive oxygen species (ROS) were evaluated by the corresponding kit. Mitochondrial membrane potential was assessed by JC-1 kit. HE staining, TUNEL and immunohistochemistry were used to detect the xenografted tumors. Protein expression was estimated by Western blot. Bupivacaine significantly inhibited the activity of T24 cells and 5637 cells at 0.25–16 mM. Bupivacaine promoted cell apoptosis with increased concentration. bupivacaine inhibited the expression of Bcl-2 and increased the expression of Bax and cytochrome C. Moreover, bupivacaine amplified the level of Fe²⁺ and ROS, and restrained the expression of cystine/glutamic acid reverse transporter (xCT) and glutathione peroxidase 4 (GPX4). Further results showed that bupivacaine decreased mitochondrial membrane potential, reduced GSH, and increased MDA levels. Besides, bupivacaine attenuated the phosphorylation of PI3K, Akt, and mTOR. In addition, bupivacaine suppressed the growth of xenografted tumors, induced apoptosis and ferroptosis, and inhibited the activity of PI3K/AKT signaling pathway in xenografted tumors. Bupivacaine could induce apoptosis and ferroptosis by inhibiting PI3K/Akt signaling pathway in bladder cancer cells.

Graphical abstract

KEYWORDS:

• Bupivacaine
• bladder cancer
• apoptosis
• ferroptosis
• PI3K/AKT signaling pathway

Highlights

• Bupivacaine inhibited growth of bladder cancer cell in vitro and vivo.

• Bupivacaine induced ferroptosis through promoting the increase of Fe²⁺ and ROS and reducing the expression of xCT and GPX4.

• Inactivity of PI3K/AKT signaling pathway might be the main molecular mechanism of bupivacaine against bladder cancer.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Author contributions

Zeqing Huang was for concept, design, and critical Review; Jianli Hao was for design, supervision, and writing Manuscript; Jianli Hao and Weiqing Zhang were for materials, data collection and processing; Jianli Hao and Weiqing Zhang were for analysis, interpretation, and literature Search.

Additional information

Funding

The author(s) reported there is no funding associated with the work featured in this article.