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Endothelin-1 Promotes Osteosarcoma Cell Invasion and Survival against Cisplatin-induced Apoptosis

  • Basic Research
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Clinical Orthopaedics and Related Research®

Abstract

Background

Endothelin-1 (ET-1) participates in a wide range of cancer-relevant processes including cell proliferation, inhibition of apoptosis, matrix remodeling, bone deposition, and metastases. Although ET-1 reportedly promotes osteosarcoma (OS) cell invasion, suggesting an important role of ET-1 in OS metastasis, the role of ET-1 in OS remains unclear.

Question/purposes

We asked whether (1) ET-1 expression is associated with the malignancy of OS, (2) ET-1 enhances the cell invasion ability of OS, and (3) ET-1 promotes OS cell survival against apoptotic stress.

Methods

We cultured primary OS specimens from 22 patients with Stages II (OS-II) and III (OS-III) in real-time quantitative RT-PCR and ELISA to compare ET-1 expression. We used Transwell® cell invasion assays (in triplicate) to assess the invasion ability of cells in the presence or absence of exogenous ET-1 and/or ET receptor antagonists. We compared cell apoptosis rate among the cells treated with cisplatin in the presence or absence of exogenous ET-1 and/or ET receptor antagonists. We used OS cell line MG-63 in all experiments as a reference.

Results

Real-time quantitative RT-PCR and ELISA showed OS-III cells had greater ET-1 expression than OS-II cells at the mRNA and the secreted protein levels. Transwell® cell invasion assays showed OS-III cells had a greater migrated cell number than OS-II cells, which could be abrogated by ETA receptor antagonist BQ123 (100 pmol/L), but not ETB receptor antagonist BQ788 (1 μmol/L); exogenous ET-1 dose-dependently promoted OS cell migration, which could be inhibited by BQ123 (100 pmol/L). Cisplatin (10 nmol/L) induced less apoptosis in OS-III cells than in OS-II cells; exogenous ET-1 dose-dependently promoted OS cell survival against cisplatin-induced apoptosis; both effects were reversed by BQ123 (1 μmol/L), but not BQ788 (1 μmol/L).

Conclusions

Increased ET-1 expression appears to be associated with increased malignancy of OS. ET-1 promotes OS cell invasion and survival against cisplatin-induced apoptosis through the ETA receptor.

Clinical Relevance

The ET-1/ETA pathway may represent an important target for treating OS, because blocking the ETA receptor with a selective antagonist can inhibit OS cell invasion and potentiate a chemotherapeutic agent’s effect on OS.

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Acknowledgments

We thank Drs. Hong Yu and Lin Chen (Department of Pathology, **angya School of Medicine, Changsha, China) for technical support on identification of tumor cells in primary osteosarcoma cell cultures. We thank Prof. Bin Shan (Department of Medicine, Tulane University, LA, USA) for help with preparation of the manuscript.

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Authors and Affiliations

  1. Department of Orthopedics, **angya Hospital, Central South University, 87 **angya Road, Changsha, Hunan, 410078, People’s Republic of China

    Yuanting Zhao MD, Qiande Liao MD, Yong Zhu MD & Haitao Long MD

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  1. Yuanting Zhao MD
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  2. Qiande Liao MD
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  3. Yong Zhu MD
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Correspondence to Qiande Liao MD.

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Each author certifies that he or she has no commercial associations (eg, consultancies, stock ownership, equity interest, patent/licensing arrangements, etc.) that might pose a conflict of interest in connection with the submitted article.

Each author certifies that his or her institution approved the human protocol for this investigation, that all investigations were conducted in conformity with ethical principles of research, and that informed consent for participation in the study was obtained.

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Zhao, Y., Liao, Q., Zhu, Y. et al. Endothelin-1 Promotes Osteosarcoma Cell Invasion and Survival against Cisplatin-induced Apoptosis. Clin Orthop Relat Res 469, 3190–3199 (2011). https://doi.org/10.1007/s11999-011-1939-2

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  • DOI: https://doi.org/10.1007/s11999-011-1939-2

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