biostudies-literatureMa XPengcheng Yingcai Medical Youth Reserve Talent Training ProjectApplied Basic Research of Yunnan Science &amp; Technology AgencyNational Natural Science Foundation of ChinaNatural Science Foundation of Jiangsu Province2493556https://www.ebi.ac.uk/biostudies/studies/S-EPMC12010658biostudies-literatureUnknown30(1)<h4>Background</h4>Pulmonary metastases in osteosarcoma (OS) are associated with a poor prognosis. Rotenone has shown anti-cancer activity. However, its effects on metastasis and the underlying mechanisms remain unknown. This study investigated the potential use of Rotenone for OS treatment.<h4>Methods</h4>The effect of Rotenone and ROS/Ca²⁺/AMPK/ZO-2 pathway on metastasis and EMT was evaluated by Western blot, Transwell and Wound healing. Flow cytometer was employed to measure the intracellular Ros and Ca²⁺ levels. The subcellular location of ZO-2 was detected by IF, interaction between AMPK and ZO-2 were examined by Co-IP. Then, subcutaneous tumor and metastasis models were used to evaluate the function of Rotenone in OS metastasis.<h4>Results</h4>Rotenone-induced RRedox report : communications in free radical researchRotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca&lt;sup&gt;2+&lt;/sup&gt;/AMPK pathway.PMC12010658BK20220235XWRCHT2022002882203396BK20231164202201AY070001[-152]Jiang KYang ZLi ZZhu ZMa XMa HWang WYunqing WChen BWang JDong SfalseRotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca&lt;sup&gt;2+&lt;/sup&gt;/AMPK pathway.<h4>Background</h4>Pulmonary metastases in osteosarcoma (OS) are associated with a poor prognosis. Rotenone has shown anti-cancer activity. However, its effects on metastasis and the underlying mechanisms remain unknown. This study investigated the potential use of Rotenone for OS treatment.<h4>Methods</h4>The effect of Rotenone and ROS/Ca²⁺/AMPK/ZO-2 pathway on metastasis and EMT was evaluated by Western blot, Transwell and Wound healing. Flow cytometer was employed to measure the intracellular Ros and Ca²⁺ levels. The subcellular location of ZO-2 was detected by IF, interaction between AMPK and ZO-2 were examined by Co-IP. Then, subcutaneous tumor and metastasis models were used to evaluate the function of Rotenone in OS metastasis.<h4>Results</h4>Rotenone-induced R2025-01-01T00:00:00Z2025 Dec2025-07-01T03:04:54.776Z2025-07-01T03:04:54.776ZS-EPMC120106584024763510.1080/13510002.2025.2493556