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Breast cancer cells expressing cancer-associated sialyl-Tn antigen have less capacity to develop osteolytic lesions in a mouse model of skeletal colonization

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Abstract

Breast cancer is one of the most prevalent malignancies in women, and approximately 75–80% of patients with advanced breast cancer develop bone metastasis. Expression of the cancer-associated carbohydrate antigen sialyl-Tn (STn) in breast cancer is associated with a poor prognosis; however, involvement of STn in the development of metastatic bone lesions remains unclear. We investigated whether STn expression on breast cancer cells influences intraosseous tumor growth and bone response in mice models of skeletal colonization. STn-positive (STn+) breast cancer cells were generated by stable transfection of an expression vector encoding ST6GaLNAc I into the breast cancer cell line MDA-MB-231. Parental MDA-MB-231 cells not expressing STn antigen were used as STn-negative (STn−) breast cancer cells. Contrary to expectations, STn expression attenuated the development of destructive bone lesions in the in vivo mice models. An in vitro study demonstrated that STn expression impaired adhesion of MDA-MB-231cells to bone marrow stromal cells. This finding in vitro was also confirmed by another breast cancer cell line MCF-7. Cell adhesion to fibronectin and type I collagen was also impaired in STn+ MDA-MB-231 cells compared to that in STn− MDA-MB-231 cells, suggesting integrin dysfunction. Given that the integrin β1 subunit is the main carrier of the STn epitope, it is likely that changes in glycan structure impaired the adhesive capacity of β1 integrin in the bone environment, leading to attenuation of tumor cell engraftment. In conclusion, breast cancer cells expressing STn antigen had less capacity for skeletal colonization, possibly due to impaired adhesive capability.

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Acknowledgements

This work was supported by the Japan Society for the Promotion of Science (JSPS) Grants-in-Aid for Scientific Research (KAKENHI) Grant Number JP15K10427 and Japan Orthopaedics and Traumatology Foundation, Inc. Grant Number 330.

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  1. Ryo Fujita and Hiroki Hamano have contributed equally to this article.

Authors and Affiliations

  1. Department of Orthopedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Japan

    Ryo Fujita, Hiroki Hamano, Yusuke Kameda, Ryuta Arai, Tomohiro Shimizu, Masahiro Ota, Dai Sato, Hideyuki Kobayashi, Norimasa Iwasaki & Masahiko Takahata

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  1. Ryo Fujita
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  2. Hiroki Hamano
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Correspondence to Masahiko Takahata.

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All animal studies were performed in accordance with protocols approved by the Hokkaido University Committee on Animal Resources.

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10585_2019_9999_MOESM1_ESM.jpg

Supplementary material 1—Immunocytochemistry and gene expression analysis to confirm the expression of STn on MCF-7 breast cancer cells. (a) Immunofluorescence microscopy confirmed that ST6GalNAc I -expressing cells expressed the STn antigen on the cell surface of MCF-7 cells. (b) STn detection by flow cytometry. The presence of the STn antigen was revealed using STn mAb (black lines) and negative controls (dotted lines, secondary antibodies only). (c) Expression of the ST6GalNAc I gene was examined by quantitative PCR. Abbreviations: STn, sialyl-Tn; mAb, monoclonal antibody. RNAi, ST6GalNAc I knockdown clones from STn+ MCF-7 cells.(JPEG 781 kb)

Supplementary material 2 (DOCX 13 kb)

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Fujita, R., Hamano, H., Kameda, Y. et al. Breast cancer cells expressing cancer-associated sialyl-Tn antigen have less capacity to develop osteolytic lesions in a mouse model of skeletal colonization. Clin Exp Metastasis 36, 539–549 (2019). https://doi.org/10.1007/s10585-019-09999-6

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  • DOI: https://doi.org/10.1007/s10585-019-09999-6

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