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Updated: May 11, 2025
A Syngeneic Orthotopic Osteosarcoma Sprague Dawley Rat Model with Amputation to Control Metastasis Rate
Published on: May 3, 2021
Rotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca2+/AMPK pathway
Xiang Ma1, Zhen Li2, Hengwei Ma3
1Department of Orthopaedics, The Third Affiliated Hospital of Kunming Medical University, Kunming, People's Republic of China.
Background:
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.
Methods:
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.
Results:
Rotenone-induced ROS led to increased intracellular Ca2+, which promoted the EMT of OS cells through activation of AMPK and ZO-2 nuclear translocation. Inhibition of ROS production decreased intracellular Ca2+, restraining AMPK activity. Knock-down of ZO-2 significantly suppressed the anti-metastasis effects of Rotenone in OS cells. Moreover, Rotenone elevated p-AMPK and ZO-2 expression but inhibited EMT and lung metastasis in vivo.Conclusion These results provide evidence supporting an anti-metastatic effect of Rotenone. These findings support the use of Rotenone in the prevention of OS metastasis.
Insights
Rotenone inhibits osteosarcoma (OS) metastasis by reducing reactive oxygen species (ROS) and calcium levels. This mechanism involves the ROS/Ca2+/AMPK/ZO-2 pathway, highlighting Rotenone
Area of Science:
- Oncology
- Molecular Biology
- Cancer Metastasis Research
Background:
- Pulmonary metastases in osteosarcoma (OS) significantly worsen patient prognosis.
- Rotenone exhibits anti-cancer properties, but its role in OS metastasis and mechanisms are unexplored.
- This study investigates Rotenone's potential therapeutic application against OS metastasis.
Purpose of the Study:
- To elucidate the anti-metastatic effects of Rotenone in osteosarcoma.
- To investigate the underlying molecular mechanisms involving the ROS/Ca2+/AMPK/ZO-2 pathway.
- To evaluate Rotenone's efficacy in preclinical OS metastasis models.
Main Methods:
- Western blot, Transwell, and wound healing assays assessed metastasis and epithelial-mesenchymal transition (EMT).
- Flow cytometry measured intracellular ROS and Ca2+ levels.
- Immunofluorescence (IF) and co-immunoprecipitation (Co-IP) examined ZO-2 localization and AMPK interaction.
Main Results:
- Rotenone-induced ROS increased intracellular Ca2+, activating AMPK and ZO-2 nuclear translocation, promoting EMT.
- Inhibition of ROS production reversed these effects, suppressing AMPK activity.
- ZO-2 knockdown diminished Rotenone's anti-metastatic effects, while Rotenone elevated p-AMPK and ZO-2, inhibiting EMT and lung metastasis *in vivo*.
Conclusions:
- Rotenone demonstrates significant anti-metastatic effects in osteosarcoma.
- The ROS/Ca2+/AMPK/ZO-2 pathway mediates Rotenone's action against OS metastasis.
- Rotenone shows promise for preventing osteosarcoma metastasis.
