Cellular and molecular effects of rocuronium and its interaction with doxorubicin in SAOS-2 osteosarcoma cells: an in vitro and in silico study

Emerging evidence suggests that anesthetic agents administered during the perioperative period may modulate tumor biology and influence chemotherapeutic efficacy. This study aimed to investigate the molecular and cellular effects of the neuromuscular blocking agents rocuronium and sugammadex on osteosarcoma cells and to evaluate their potential interaction with doxorubicin, using integrated in vitro and in silico approaches. SAOS-2 osteosarcoma cells were treated with rocuronium, sugammadex, and doxorubicin alone or in fixed-ratio combinations. Cell viability and drug interactions were evaluated by MTT assay and Chou–Talalay analysis. Cell migration was assessed using scratch assays, apoptosis- and migration-related gene expression by qRT-PCR, and molecular docking was performed to investigate interactions of rocuronium and doxorubicin with HSP70, HSP90, and CYP450. Rocuronium and sugammadex exhibited modest antiproliferative effects compared with doxorubicin; however, rocuronium demonstrated greater biological activity and was selected for further analyses. Combined treatment with rocuronium and doxorubicin showed a dose-dependent synergistic antiproliferative interaction at higher fractional effect levels, accompanied by favorable dose-reduction indices for doxorubicin. Both agents significantly inhibited SAOS-2 cell migration when applied individually. Gene expression analyses revealed coordinated activation of apoptotic pathways, with doxorubicin predominantly engaging intrinsic, mitochondria-dependent apoptosis, while rocuronium additionally activated extrinsic caspase-8–mediated signaling. Molecular docking predicted preferential binding of doxorubicin to HSP70 and HSP90, whereas rocuronium showed the strongest interaction with CYP450, suggesting complementary molecular targeting profiles. This study suggests that rocuronium may modulate the response of osteosarcoma cells to doxorubicin under in vitro conditions, potentially involving changes in apoptosis- and stress-related molecular pathways. These findings support further investigation of whether perioperative exposure to neuromuscular blocking agents may influence chemotherapy-related cellular responses in osteosarcoma.