Background: Gallbladder cancer (GBC) is a highly aggressive malignancy with a notoriously poor prognosis and limited treatment options. Intriguingly, beyond its established perioperative applications, the inhalational anesthetic Sevoflurane has garnered attention for its potential anti-tumor effects, demonstrating the ability to inhibit progression in various cancers. However, its specific impact and the underlying molecular mechanisms in GBC remain largely unexplored. This study systematically investigated the anti-cancer property of Sevoflurane in GBC and associated signaling pathways.

Methods: Sevoflurane at 1.7%, 3.4% and 5.1% concentrations was separately employed to treat human intestinal biliary epithelial cells (HIBEpic) and GBC-SD cells. GBC-SD cells were also pretreated with an endoplasmic reticulum (ER) stress inhibitor Salubrinal, or transfected with Rho-associated coiled-coil containing protein kinase 1 (ROCK1)-overexpressing vectors/short hairpin RNA against ROCK1. Cell viability, migration and invasion, colony formation, and apoptosis were determined using Cell Counting Kit-8, Transwell, colony formation, and flow cytometry assays, respectively. Special kits were used to detect levels of glucose uptake, lactate and adenosine triphosphate (ATP) in GBC-SD cells. ROCK1 expression and levels of ROCK1, activating transcription factor 4 (ATF4) and C/EBP-homologous protein 10 (CHOP) were quantified by quantitative real-time reverse transcription polymerase chain reaction and Western blot analyses, respectively.

Results: Sevoflurane exposure inhibited malignant phenotypes, promoted apoptosis and expressions of ATF4 and CHOP, and reduced ROCK1 expression and levels of glucose uptake, lactate and ATP in GBC-SD cells, while Salubrinal pretreatment reversed the upregulated trends of ATF4 and CHOP (p < 0.05). ROCK1 was highly expressed in different GBC cell lines (p < 0.05). ROCK1 overexpression enhanced malignant phenotypes, promoted levels of glucose uptake, lactate and ATP, and suppressed apoptosis in GBC-SD cells, while ROCK1 silencing reversed these alterations (p < 0.05). Meanwhile, ROCK1 overexpression reversed the effects of Sevoflurane on GBC-SD cells (p < 0.05).

Conclusion: Sevoflurane suppresses GBC progression by activating ER stress and reducing glycolysis via ROCK1 downregulation.