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Insulin-like growth factor binding protein 5 suppresses tumor growth and metastasis of human osteosarcoma

Oncogene volume 30, pages 3907–3917 (2011)Cite this article

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Abstract

Osteosarcoma (OS) is the most common primary malignancy of bone. There is a critical need to identify the events that lead to the poorly understood mechanism of OS development and metastasis. The goal of this investigation is to identify and characterize a novel marker of OS progression. We have established and characterized a highly metastatic OS subline that is derived from the less metastatic human MG63 line through serial passages in nude mice via intratibial injections. Microarray analysis of the parental MG63, the highly metastatic MG63.2 subline, as well as the corresponding primary tumors and pulmonary metastases revealed insulin-like growth factor binding protein 5 (IGFBP5) to be one of the significantly downregulated genes in the metastatic subline. Confirmatory quantitative RT–PCR on 20 genes of interest demonstrated IGFBP5 to be the most differentially expressed and was therefore chosen to be one of the genes for further investigation. Adenoviral mediated overexpression and knockdown of IGFBP5 in the MG63 and MG63.2 cell lines, as well as other OS lines (143B and MNNG/HOS) that are independent of our MG63 lines, were employed to examine the role of IGFBP5. We found that overexpression of IGFBP5 inhibited in vitro cell proliferation, migration and invasion of OS cells. Additionally, IGFBP5 overexpression promoted apoptosis and cell cycle arrest in the G1 phase. In an orthotopic xenograft animal model, overexpression of IGFBP5 inhibited OS tumor growth and pulmonary metastases. Conversely, siRNA-mediated knockdown of IGFBP5 promoted OS tumor growth and pulmonary metastases in vivo. Immunohistochemical staining of patient-matched primary and metastatic OS samples demonstrated decreased IGFBP5 expression in the metastases. These results suggest 1) a role for IGFBP5 as a novel marker that has an important role in the pathogenesis of OS, and 2) that the loss of IGFBP5 function may contribute to more metastatic phenotypes in OS.

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Acknowledgements

We thank Dr Xinmin Li, PhD (Director of The UCLA Clinical Microarray Core) for his help with the microarray and cluster analysis. We also sincerely thank Dr Anthony Montag, Leslie Martin and Guarav Luther for the creation of the tissue array. We also thank Dr Theodore Karrison for help with the statistical analysis. This work was supported in part by research grants from The American Cancer Society (HHL and TCH), the Brinson Foundation (HHL, TCH, RCH), The National Institute of Health (HHL, TCH, RCH), the Orthopaedic Research and Education Foundation (HHL, RCH), the Natural Science Foundation of China (#81001197 to YS), and the 973 Program of the Ministry of Science and Technology of China (#2011CB707906 to JL and TCH).

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  1. Y Su and E R Wagner: These authors contributed equally to this work.

Authors and Affiliations

  1. Stem Cell Biology and Therapy Laboratory, The Children's Hospital of Chongqing Medical University, Chongqing, China

    Y Su, Q Luo, G Zhu, Y Bi, M Li, J Qin & T-C He

  2. Department of Surgery, The University of Chicago Medical Center, Chicago, IL, USA

    Y Su, E R Wagner, Q Luo, J Huang, L Chen, B-C He, G-W Zuo, Q Shi, B-Q Zhang, G Zhu, Y Bi, J Luo, X Luo, S H Kim, J Shen, F Rastegar, E Huang, Y Gao, J-L Gao, K Yang, J Qin, R C Haydon, T-C He & H H Luu

  3. Key Laboratory of Diagnostic Medicine Designated by Chinese Ministry of Education and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China

    Y Su, Q Luo, J Huang, L Chen, B-C He, G-W Zuo, Q Shi, B-Q Zhang, G Zhu, Y Bi, J Luo, X Luo, M Li, J Qin & T-C He

  4. School of Bioengineering, Chongqing University, Chongqing, China

    E Huang

  5. Department of Geriatrics, Xinhua Hospital of Shanghai Jiatong University, Shanghai, China

    Y Gao

  6. Department of Cell Biology, The Third Military Medical University, Chongqing, China

    K Yang

  7. Department of Radiology, The University of Chicago Medical Center, Chicago, IL, USA

    C Wietholt

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  1. Y Su
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Correspondence to H H Luu.

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Su, Y., Wagner, E., Luo, Q. et al. Insulin-like growth factor binding protein 5 suppresses tumor growth and metastasis of human osteosarcoma. Oncogene 30, 3907–3917 (2011). https://doi.org/10.1038/onc.2011.97

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