Masato Yoshizawa1, Seikou Nakamura2, Yuki Sugiyama1, Shiori Tamai1, Yukiko Ishida1, Mari Sueyoshi1, Yuki Toda1, Shigekuni Hosogi1, Yoshitaka Yano3, Eishi Ashihara4. 1. Department of Clinical and Translational Physiology, Kyoto Pharmaceutical University, Kyoto, Japan. 2. Department of Pharmacognosy, Kyoto Pharmaceutical University, Kyoto, Japan. 3. Education and Research Center for Clinical Pharmacy, Kyoto Pharmaceutical University, Kyoto, Japan. 4. Department of Clinical and Translational Physiology, Kyoto Pharmaceutical University, Kyoto, Japan ash@mb.kyoto-phu.ac.jp.
Abstract
BACKGROUND/AIM: Osteosarcoma is the most malignant type of bone tumor. Patients with osteosarcoma metastases have a poorer prognosis than those without metastases. Thus, the prognosis of osteosarcoma patients with metastases must be improved. MATERIALS AND METHODS: The present study investigated the inhibitory effects of 6-hydroxythiobinupharidine isolated from Nuphar pumilum on migration of LM8 murine osteosarcoma cells by a migration assay and also examined the expression of proteins related to actin dynamics by western blot. The present study also developed an automatic cell counting system using machine learning to count migrated cells by Fiji and Trainable Weka Segmentation. RESULTS: 6-Hydroxythiobinupharidine inhibited migration of LM8 osteosarcoma cells in a dose-dependent manner, and decreased protein expression of Lin11, Isl-1, and Mec-3 domain kinase 1 (LIMK1) and the levels of phosphorylated Cofilin. CONCLUSION: 6-Hydroxythiobinupharidine suppressed migration of LM8 osteosarcoma cells by decreasing expression of LIMK1. 6-Hydroxythiobinupharidine could be potentially used as an anti-metastatic compound. Copyright
BACKGROUND/AIM: Osteosarcoma is the most malignant type of bone tumor. Patients with osteosarcoma metastases have a poorer prognosis than those without metastases. Thus, the prognosis of osteosarcomapatients with metastases must be improved. MATERIALS AND METHODS: The present study investigated the inhibitory effects of 6-hydroxythiobinupharidine isolated from Nuphar pumilum on migration of LM8murineosteosarcoma cells by a migration assay and also examined the expression of proteins related to actin dynamics by western blot. The present study also developed an automatic cell counting system using machine learning to count migrated cells by Fiji and Trainable Weka Segmentation. RESULTS:6-Hydroxythiobinupharidine inhibited migration of LM8osteosarcoma cells in a dose-dependent manner, and decreased protein expression of Lin11, Isl-1, and Mec-3 domain kinase 1 (LIMK1) and the levels of phosphorylated Cofilin. CONCLUSION:6-Hydroxythiobinupharidine suppressed migration of LM8osteosarcoma cells by decreasing expression of LIMK1. 6-Hydroxythiobinupharidine could be potentially used as an anti-metastatic compound. Copyright