Enhancing intracellular reactive oxygen species, apoptosis, and wound healing activities of Benjakul in cholangiocarcinoma

Introduction: Cholangiocarcinoma (CCA) is an aggressive malignancy of the biliary tract with limited treatment options and a poor prognosis. Current therapies, such as gemcitabine and cisplatin, are associated with notable side effects and limited efficacy. Benjakul (BJK), a traditional Thai herbal formulation, has emerged as a potential anticancer agent. The present study aimed to assess the anticancer effects of BJK on CCA cell lines during cancer progression, focusing on its ability to induce oxidative stress, promote apoptosis and inhibit cell migration. Methods: BJK extract at concentrations of ۵۰, ۱۰۰ and ۲۰۰ µg/mL was applied to CCA, KKU-۲۱۳B, and KKU-۱۰۰ cell lines. Intracellular reactive oxygen species (ROS) and apoptosis levels were measured using flow cytometry, whilst cell migration was assessed using a wound-healing assay. Results: BJK extract increased intracellular ROS levels, particularly in KKU-۲۱۳B cells, leading to apoptosis. At the concentrations of ۵۰, ۱۰۰, and ۲۰۰ µg/mL, BJK significantly induced apoptosis in KKU-۲۱۳B cells at the rates of ۱۶.۲, ۵۱.۷ and ۵۶.۷% (P<۰.۰۵), respectively, whilst its effects on KKU-۱۰۰ cells were minimal. Furthermore, BJK demonstrated potent anti-migratory effects, significantly inhibiting the migration of KKU-۲۱۳B cells in the wound-healing assay (P<۰.۰۱). Conclusion: BJK may exert its anticancer effects through intracellular ROS-mediated apoptosis and migration inhibition, with the combination of its constituent herbs enhancing its therapeutic potential. Furthermore, the results highlight the potential of BJK as a promising therapeutic agent for CCA. By leveraging the anticancer properties of traditional herbal formulations, BJK may improve treatment outcomes for patients with this challenging disease.