Berberine Chloride Suppresses Growth of Canine Osteosarcoma Cells via Regulating MMP2/AKT/β-Catenin Pathway.

Canine osteosarcoma is an aggressive primary bone malignancy associated with a poor prognosis and a high propensity for lung metastasis. Due to the limitations and side effects of conventional chemotherapy, there remains a pressing need for alternative therapeutic strategies and adjuvants. In this study, we examined the effects of berberine chloride (BBRC), a bioactive isoquinoline alkaloid derived from the dried roots of Coptidis Rhizoma, on canine osteosarcoma D-17 and DSN cell lines. BBRC reduced the viability and migratory capacity of D-17 and DSN cell lines in three-dimensional (3D) spheroid cultures. Additionally, BBRC induced calcium ion overload in the mitochondrial matrix but not in the cytoplasm, in both osteosarcoma cell lines. BBRC also triggered excessive generation of reactive oxygen species (ROS) in both cell lines. Furthermore, Western Blot analysis revealed that BBRC downregulated the MMP2/AKT/β-catenin signaling pathways in both BBRC in D-17 and DSN cells. In addition, BBRC downregulated Cyclin D1 and Survivin, known downstream targets of the β-catenin pathway. We also confirmed that BBRC suppressed the transcription of genes involved in cell cycle regulation. Overall, our findings indicate that BBRC inhibits osteosarcoma cell growth by modulating calcium-redox homeostasis and the MMP2/AKT/β-catenin pathway.