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Interstitial Fluid Sphingosine-1-Phosphate in Murine Mammary Gland and Cancer and Human Breast Tissue and Cancer Determined by Novel Methods

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Abstract

The tumor microenvironment is a determining factor for cancer biology and progression. Sphingosine-1-phosphate (S1P), produced by sphingosine kinases (SphKs), is a bioactive lipid mediator that regulates processes important for cancer progression. Despite its critical roles, the levels of S1P in interstitial fluid (IF), an important component of the tumor microenvironment, have never previously been measured due to a lack of efficient methods for collecting and quantifying IF. The purpose of this study is to clarify the levels of S1P in the IF from murine mammary glands and its tumors utilizing our novel methods. We developed an improved centrifugation method to collect IF. Sphingolipids in IF, blood, and tissue samples were measured by mass spectrometry. In mice with a deletion of SphK1, but not SphK2, levels of S1P in IF from the mammary glands were greatly attenuated. Levels of S1P in IF from mammary tumors were reduced when tumor growth was suppressed by oral administration of FTY720/fingolimod. Importantly, sphingosine, dihydro-sphingosine, and S1P levels, but not dihydro-S1P, were significantly higher in human breast tumor tissue IF than in the normal breast tissue IF. To our knowledge, this is the first reported S1P IF measurement in murine normal mammary glands and mammary tumors, as well as in human patients with breast cancer. S1P tumor IF measurement illuminates new aspects of the role of S1P in the tumor microenvironment.

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Acknowledgments

We gratefully acknowledge the VCU Lipidomics Core, which is supported in part by funding from the NIH-NCI Cancer Center Support Grant P30CA016059.

Author Contribution

MN and KT conceived the study. MN, AY, TA, WCH, KPT, BA, carried out experiments. HM developed the IF collection tube. JCA performed mass spectrometry analysis. MN and TK wrote the manuscript with assistance from OMR, TW, SS and SM.

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Authors and Affiliations

  1. Division of Surgical Oncology, Department of Surgery, Virginia Commonwealth University School of Medicine, and Massey Cancer Center, PO Box 980011, West Hospital 7-402, 1200 East Broad Street, Richmond, VA, 23298-0011, USA

    Masayuki Nagahashi, Akimitsu Yamada, Tomoyoshi Aoyagi, Wei-Ching Huang, Krista P. Terracina, Barbara J. Adams & Kazuaki Takabe

  2. Department of Biochemistry and Molecular Biology, Virginia Commonwealth University School of Medicine, and the Massey Cancer Center, PO Box 980011, West Hospital 7-402, 1200 East Marshall Street, Richmond, VA, 23298, USA

    Masayuki Nagahashi, Akimitsu Yamada, Jeremy C. Allegood, Tomoyoshi Aoyagi, Wei-Ching Huang, Krista P. Terracina, Barbara J. Adams, Sheldon Milstien, Sarah Spiegel & Kazuaki Takabe

  3. Division of Digestive and General Surgery, Niigata University Graduate School of Medical and Dental Sciences, 1-757 Asahimachi-dori, Chuo-ku, Niigata City, Niigata, 951-8510, Japan

    Masayuki Nagahashi, Junko Tsuchida & Toshifumi Wakai

  4. Section of General Internal Medicine, Kojin Hospital, 1-710 Shikenya, Moriyama, Nagoya, 463-8530, Japan

    Hiroshi Miyazaki

  5. Holy Cross Hospital Michael and Dianne Bienes Comprehensive Cancer Center, 4725 North Federal Highway, Fort Lauderdale, FL, 33308, USA

    Omar M. Rashid

  6. Massachusetts General Hospital, 55 Fruit St, Boston, MA, 02114, USA

    Omar M. Rashid

  7. University of Miami Miller School of Medicine, 1600 NW 10th Ave, Miami, FL, 33136, USA

    Omar M. Rashid

  8. Breast Surgery, Roswell Park Cancer Institute, Elm & Carlton Streets, Buffalo, NY, 14263, USA

    Kazuaki Takabe

Authors
  1. Masayuki Nagahashi
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  2. Akimitsu Yamada
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  3. Hiroshi Miyazaki
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  4. Jeremy C. Allegood
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  5. Junko Tsuchida
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  6. Tomoyoshi Aoyagi
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  7. Wei-Ching Huang
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  8. Krista P. Terracina
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  9. Barbara J. Adams
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  10. Omar M. Rashid
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  11. Sheldon Milstien
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  12. Toshifumi Wakai
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  13. Sarah Spiegel
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  14. Kazuaki Takabe
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Corresponding authors

Correspondence to Masayuki Nagahashi or Kazuaki Takabe.

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The authors declare no conflict of interest.

Funding

MN was a Japan Society for the Promotion of Science Postdoctoral Fellow. MN is supported by the Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research Grant Number 15H05676 and 15 K15471, the Uehara Memorial Foundation, Nakayama Cancer Research Institute, Takeda Science Foundation, and Tsukada Medical Foundation. JT is supported by the JSPS Grant-in-Aid for Scientific Research Grant Number 16 K19888. KT is supported by NIH/NCI grant R01CA160688 and Susan G. Komen Investigator Initiated Research Grant IIR12222224. SS is supported by NIH/NCI grant R01CA61774 and in part by the DoD BCRP program under award number W81XWH-14-1-0086. KPT is supported by T32CA085159.

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Nagahashi, M., Yamada, A., Miyazaki, H. et al. Interstitial Fluid Sphingosine-1-Phosphate in Murine Mammary Gland and Cancer and Human Breast Tissue and Cancer Determined by Novel Methods. J Mammary Gland Biol Neoplasia 21, 9–17 (2016). https://doi.org/10.1007/s10911-016-9354-7

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  • DOI: https://doi.org/10.1007/s10911-016-9354-7

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