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Updated: Dec 17, 2025
LINE-1 Methylation Analysis in Mesenchymal Stem Cells Treated with Osteosarcoma-Derived Extracellular Vesicles
Published on: February 1, 2020
Untargeted LC-MS/MS analysis reveals metabolomics feature of osteosarcoma stem cell response to methotrexate
Feng Wang1, Zhiyu Zhang1, Qin Li2
1Department of Orthopedics, the Fourth Affiliated Hospital of China Medical University, Chongshan Road, Shenyang, 110032 Liaoning People's Republic of China.
Background:
Cancer stem cell (CSC) is identified in osteosarcoma (OS) and considered resistant to chemotherapeutic agents. However, the mechanism of osteosarcoma stem cell (OSC) resistant to chemotherapy remains debatable and vague, and the metabolomics feature of OSC is not clarified.
Materials And Methods:
OSC was isolated by using sphere forming assay and identified. Untargeted LC-MS/MS analysis was performed to reveal the metabolomics feature of OSC and underlying mechanisms of OSC resistant to methotrexate (MTX).
Results:
OSC was efficiently isolated and identified from human OS 143B and MG63 cell lines with enhanced chemo-resistance to MTX. The untargeted LC-MS analysis revealed that OSC showed differential metabolites and perturbed signaling pathways, mainly involved in metabolisms of fatty acid, amino acid, carbohydrate metabolism and nucleic acid. After treated with MTX, metabolomics feature of OSC was mainly involved metabolisms of amino acid, fatty acid, energy and nucleic acid. Moreover, compared with their parental OS cells response to MTX, the differential metabolites and perturbed signaling pathways were mainly involved in metabolism of amino acid, fatty acid and nucleic acid. What's more, Rap1 signaling pathway and Ras signaling pathway were involved in OS cells and their SCs response to MTX.
Conclusion:
Sphere-forming assay was able to efficiently isolate OSC from human OS cell lines and the untargeted LC-MS/MS analysis was suggested a sufficient methodology to investigate metabolomics features of OS cells and OSCs. Moreover, the metabolomics features of OSCs response to MTX might reveal a further understanding of chemotherapeutic resistance in OS.
Insights
Osteosaroma stem cells (OSCs) show enhanced resistance to methotrexate (MTX). Metabolomics analysis reveals altered metabolic pathways in OSCs, offering insights into chemotherapy resistance mechanisms in osteosarcoma.
Area of Science:
- Oncology
- Stem Cell Biology
- Metabolomics
Background:
- Cancer stem cells (CSCs) are implicated in osteosarcoma (OS) chemoresistance.
- Mechanisms underlying osteosarcoma stem cell (OSC) chemoresistance are not fully understood.
- The metabolomic profile of OSCs remains largely uncharacterized.
Purpose of the Study:
- To isolate and identify osteosaroma stem cells (OSCs).
- To investigate the metabolomic features of OSCs.
- To elucidate the mechanisms of OSC resistance to methotrexate (MTX).
Main Methods:
- Osteosaroma stem cells (OSCs) were isolated using sphere-forming assays.
- Untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed for metabolomics analysis.
- Comparative analysis of OSCs and parental OS cells treated with MTX.
Main Results:
- OSCs were successfully isolated from human OS cell lines (143B and MG63) exhibiting increased MTX resistance.
- Metabolomics analysis revealed distinct metabolic profiles in OSCs, involving fatty acid, amino acid, carbohydrate, and nucleic acid metabolism.
- MTX treatment altered amino acid, fatty acid, energy, and nucleic acid metabolism in OSCs, with similar perturbations observed in parental OS cells.
- Rap1 and Ras signaling pathways were implicated in the response of OS cells and OSCs to MTX.
Conclusions:
- Sphere-forming assays are effective for isolating OSCs.
- Untargeted LC-MS/MS is a suitable method for studying OS and OSC metabolomics.
- The identified metabolomic features of OSCs provide insights into chemotherapy resistance in osteosarcoma.
