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Updated: Apr 23, 2026
LINE-1 Methylation Analysis in Mesenchymal Stem Cells Treated with Osteosarcoma-Derived Extracellular Vesicles
Published on: February 1, 2020
[Proteomics research of bufalin-induced apoptosis in osteosarcoma cell lines]
Objective:
To study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.
Method:
MTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.
Result:
Bufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.
Conclusion:
Bufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.
Insights
Bufalin effectively inhibits osteosarcoma cell growth and induces apoptosis. This effect is linked to the downregulation of heat shock protein 27 (Hsp27), a key anti-apoptotic protein.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Context:
- Osteosarcoma is a primary bone malignancy with limited treatment options.
- Bufalin, a cardiotonic steroid, has shown potential anti-cancer properties.
- Understanding bufalin's mechanism in osteosarcoma is crucial for therapeutic development.
Purpose:
- To investigate the apoptosis-inducing effects of bufalin on human osteosarcoma cells.
- To elucidate the molecular mechanisms underlying bufalin's anti-cancer activity.
- To identify specific protein targets involved in bufalin-mediated apoptosis.
Summary:
- Bufalin demonstrated dose- and time-dependent inhibition of osteosarcoma cell growth (U20S, U20S/MTX300, SAOS-2, IOR/OS9).
- Flow cytometry confirmed bufalin-induced apoptosis in these cells.
- Proteomic analysis revealed significant downregulation of heat shock protein 27 (Hsp27), an anti-apoptotic protein, which was validated by Western blot. Overexpression of Hsp27 attenuated bufalin-induced apoptosis.
Impact:
- Bufalin exhibits potent anti-proliferative and pro-apoptotic effects on human osteosarcoma cells.
- The downregulation of Hsp27 is identified as a critical mechanism in bufalin-induced apoptosis.
- Bufalin's multi-target action, particularly on Hsp27, suggests its potential as a therapeutic agent for osteosarcoma.
