Abstract
Purpose
Although undergoing conventional chemotherapy significantly improves the prognosis of Osteosarcoma, chemoresistance and failure of therapy is still a significant challenge. Furthermore, conventional chemotherapy, like doxorubicin, would upregulate the expression of programmed death-ligand 1 (PD-L1) which caused an immunosuppressive microenvironment and unsatisfied treatment result in Osteosarcoma. Thus, it is urgent to explore a strategy to overcome this disadvantage.
Methods
Human Osteosarcoma cell line MG63 and mouse Osteosarcoma cell line K7 were included in this study. Subcutaneous tumor model was used by injection of K7 cells in BALB/C mice to test the effect of doxorubicin and sorafenib on tumor growth. PD-L1 expression was tested in vitro (flow cytometry, western blot and PCR) and in vivo (flow cytometry and immunohistochemistry). Proportion of immune cells (CD4, CD8, Tregs, and cytotoxic T lymphocytes) in vivo was analyzed with flow cytometry.
Results
Combination of sorafenib and doxorubicin inhibited tumor growth significantly in vivo. Doxorubicin increased PD-L1 expression in vitro and in vivo, while sorafenib inhibited doxorubicin-induced PD-L1 upregulation in vitro and in vivo. Proportion of interferon-γ-secreting CD8 + T lymphocytes in tumor tissue was increased significantly when sorafenib was combined with doxorubicin, while proportion of CD4, CD8, and Tregs was not significantly changed. Extracellular signal-regulated kinases (ERK) pathway could be one of the key mechanisms by which doxorubicin induced upregulation of PD-L1 in Osteosarcoma cells.
Conclusion
Combination of sorafenib and conventional chemotherapeutic reagents is a potent strategy to improve treatment effectiveness by modulating tumor microenvironment in Osteosarcoma through increasing proportion of cytotoxic T lymphocytes.
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Availability of data and materials
The data generated in this study are available upon request from the corresponding author.
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Acknowledgements
Thanks to Dr Zhengdong Cai for generously supporting of mouse Osteosarcoma cell line K7. This work was supported by the National Nature Science Foundation of China (No.81702661).
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This work was supported by the National Nature Science Foundation of China (No. 81702661).
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JXW and QYB conceived and designed the experiment. JZW, FQH and PY performed the experiments and wrote the manuscript draft. JXW, JW and ZCL edited the paper. All the authors have read and approved the final manuscript and agree to be accountable for all aspects of the study.
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This study was performed in line with the principles of the Declaration of Helsinki. The protocol was approved by the animal ethical committee of Ruijin Hospital, affiliated with Shanghai Jiaotong University School of Medicine.
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Supplementary file1 Supplemental Fig. 1 Western blotting analysis of PD-L1, phosphorylation level of ERK, mTOR, and Stat3 pathways was implemented in 143b and U2 cell lines. (TIF 1827 KB)
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Wang, J., Hu, F., Yu, P. et al. Sorafenib inhibits doxorubicin-induced PD-L1 upregulation to improve immunosuppressive microenvironment in Osteosarcoma. J Cancer Res Clin Oncol 149, 5127–5138 (2023). https://doi.org/10.1007/s00432-022-04458-4
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DOI: https://doi.org/10.1007/s00432-022-04458-4