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Tumor-α9β1 integrin-mediated signaling induces breast cancer growth and lymphatic metastasis via the recruitment of cancer-associated fibroblasts

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Abstract

Tumor-derived matricellular proteins such as osteopontin (OPN) and tenascin-C (TN-C) have been implicated in tumor growth and metastasis. However, the molecular basis of how these proteins contribute to tumor progression remains to be elucidated. Importantly, these matricellular proteins are known to interact with α9β1 integrin. Therefore, we hypothesized that tumor-derived α9β1 integrin may contribute to tumor progression. To clarify the roles of α9β1 integrin in tumor growth and lymphatic metastasis, we used an inhibitory anti-human α9β1 integrin antibody (anti-hα9β1 antibody) and a α9β1 integrin-positive human breast cancer cell line, MDA-MB-231 luc-D3H2LN (D3H2LN), in vitro functional assays, and an in vivo orthotopic xenotransplantation model. In this study, we demonstrated that tumor, but not host α9β1 integrin, contributes to tumor growth, lymphatic metastasis, recruitment of cancer-associated fibroblasts (CAFs), and host-derived OPN production. We also found that CAFs contributed to tumor growth, lymphatic metastasis, and host-derived OPN levels. Consistent with those findings, tumor volume was well-correlated with numbers of CAFs and levels of host-derived OPN. Furthermore, it was shown that the inoculation of D3H2LN cells into mammary fat pads with mouse embryonic fibroblasts (MEFs), obtained from wild type, but not OPN knock-out mice, resulted in enhancement of tumor growth, thus indicating that CAF-derived OPN enhanced tumor growth. These results suggested that tumor α9β1-mediated signaling plays a pivotal role in generating unique primary tumor tissue microenvironments, which favor lymphatic metastasis and tumor growth.

Key messages

  • Tumor α9β1 integrin promotes lymphatic metastasis through enhancing invasion.

  • Tumor α9β1 integrin promotes tumor growth through CAFs.

  • Tumor α9β1 integrin enhances the recruitment of CAFs into the primary tumor.

  • Tumor cells induce the production of OPN by CAFs in the primary tumor.

  • CAF-derived OPN promotes tumor growth.

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Acknowledgments

We thank Chiemi Kimura and Orie Yamamori (Hokkaido University) for excellent technical assistance, and Tetsuro Nakamura (Gene Techono Science Corporation, Sapporo, Japan) for valuable suggestion about the use of anti-hα9β1 antibody and for valuable discussions during course of experiments. This study was supported by grant-in-aids from the Ministry of Education, Culture, Science, Sports, and Technology of Japan (21390113(B)) to Toshimitsu Uede.

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There are no potential conflicts of interest for all authors.

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Authors and Affiliations

  1. Department of Matrix Medicine, Institute for Genetic Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo, Hokkaido, 060-0815, Japan

    Daichi Ota, Yutaka Matsui, Koyu Ito & Toshimitsu Uede

  2. Division of Molecular Immunology, Institute for Genetic Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo, Hokkaido, 060-0815, Japan

    Masashi Kanayama & Naoyoshi Maeda

  3. Department of Surgery I, Sapporo Medical University Hospital, Sapporo Medical University, S-1, W-16, Chuo-Ku, Sapporo, Hokkaido, 060-8543, Japan

    Goro Kutomi & Koichi Hirata

  4. Department of Pathology I, Sapporo Medical University School of Medicine, Sapporo, Hokkaido, 060-8556, Japan

    Toshihiko Torigoe & Noriyuki Sato

  5. Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo, Hokkaido, 060-0815, Japan

    Akinori Takaoka

  6. The London Regional Cancer Program, Departments of Oncology, Pathology and Medical Biophysics, University of Western Ontario, 790 Commissioners Road East, London, ON, N6A 4 L6, Canada

    Ann F. Chambers

  7. Institute for Enzyme Research, Tokushima University, 3-18-15, Kuramoto, Tokushima, 770-8503, Japan

    Junko Morimoto

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  1. Daichi Ota
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  2. Masashi Kanayama
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  3. Yutaka Matsui
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  13. Toshimitsu Uede
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Correspondence to Daichi Ota or Toshimitsu Uede.

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Ota, D., Kanayama, M., Matsui, Y. et al. Tumor-α9β1 integrin-mediated signaling induces breast cancer growth and lymphatic metastasis via the recruitment of cancer-associated fibroblasts. J Mol Med 92, 1271–1281 (2014). https://doi.org/10.1007/s00109-014-1183-9

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  • DOI: https://doi.org/10.1007/s00109-014-1183-9

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