(271) Berberine Chloride Inhibits Growth of Canine Osteosarcoma Cells via Regulation of MMP2/AKT/

Minha Jeong¹; Juhyeong Seo¹ ; Jiyeon Ham¹

1. Department of Animal Science and Biotechnology, Chungnam National University, Daejeon, Republic of Korea

Abstract Text: Osteosarcoma is an aggressive primary bone malignancy with a poor prognosis and a high tendency to metastasize, especially to the lungs. Due to the limitations and side effects of conventional chemotherapy, there is an ongoing need to develop alternative therapeutic strategies and adjuvants to overcome these challenges. In this study, we investigated the effects of berberine chloride (BBRC), a more effective form of isoquinoline alkaloid extracted from the dried roots of Coptidis Rhizoma, on canine osteosarcoma D17 and DSN cells. We verified that BBRC inhibited the viability and migratory ability of D17 and DSN cells, including in 3D spheroid culture. Additionally, BBRC caused calcium ion overload in the mitochondrial matrix, but not in the cytoplasm, in both osteosarcoma cell lines. Notably, BBRC induced excessive reactive oxygen species (ROS) generation in both cell lines. Further, we investigated the intracellular signaling pathways via western blotting and confirmed that the MMP2/AKT/β-catenin signaling cascades were downregulated by BBRC in D17 and DSN cells. In addition, Cyclin D1 and Survivin proteins, which are downstream targets of this pathway, were also decreased by BBRC. Furthermore, we confirmed the suppression of cell cycle-related genes at the transcriptional level. Overall, our findings suggest that BBRC suppresses the cell growth of osteosarcoma via regulation of calcium-redox homeostasis and the MMP2/AKT/β-catenin pathway.