Tumor Biology

, Volume 37, Issue 4, pp 5397–5404 | Cite as

Circulating tumor-associated neutrophils (cTAN) contribute to circulating tumor cell survival by suppressing peripheral leukocyte activation

  • Juechao Zhang
  • Xuan Qiao
  • Huifang Shi
  • Xiaoqing Han
  • Wenguang Liu
  • Xiujuan Tian
  • Xianlu Zeng
Original Article


During malignant progression, primary tumors rebuild leukocyte profile and suppress the host anti-tumor immune response. Tumor-associated neutrophils (TAN) increased in the cancer patients and emerged as an important participant and regulator of immune responses. The aim of this study is to investigate the role of circulating TAN (cTAN) in the metastatic process of advanced malignancy. We tested circulating neutrophils from patients (n = 180) with various types of cancer using flow cytometry analyses. We also used B16F10 cell-implanted C57BL/6 tumor-bearing mice model to simulate the advanced malignancy. Peripheral neutrophils were isolated by ficoll density gradient centrifugation, and in vitro tumor-leukocyte co-culture model was used to test tumor cell survival under leukocyte challenge condition. Here, we showed that neutrophils increased in the peripheral blood under the pathological condition of advanced malignancy both in cancer patients and in tumor-bearing mice. In mouse model, the malignantly increased neutrophils were identified as TAN according to the gene transcriptional analyses. We also showed that cTAN enhance tumor metastasis and cTAN could inhibit the activation of the peripheral leukocytes and rescue tumor cells from leukocyte challenge. In conclusion, our finding suggests that the abundance of cTAN in advanced cancer patients contributes to the circulating tumor cell survival by suppressing peripheral leukocyte activation.


Neutrophil Tumor-associated neutrophil Tumor metastasis Tumor cell survival 



This work was supported by the grants from National Nature Science Foundation of China (81372288, 81172014) and the Natural Science Foundation of Jilin province (20150101186JC). We also would like to acknowledge the assistance of the Experimental Animal Center of NENU for the animal care, the Clinical Laboratory of the Second People’s Hospital of Jilin for the collection of clinical blood samples, and Yanyan Gao (NENU) for the stable transfected GFP positive B16F10 cell line.

Compliance of ethical standards

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration andits later amendments or comparable ethical standards.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Conflicts of interest


Supplementary material

13277_2015_4349_MOESM1_ESM.doc (75 kb)
Figure S1 (DOC 75 kb)


  1. 1.
    Kaplan RN, Psaila B, Lyden D. Bone marrow cells in the ‘pre-metastatic niche’: within bone and beyond. Cancer Metastasis Rev. 2006;25:521–9.CrossRefPubMedGoogle Scholar
  2. 2.
    Mareel MM, Van Roy FM, Bracke ME. How and when do tumor cells metastasize? Crit Rev Oncog. 1993;4:559–94.PubMedGoogle Scholar
  3. 3.
    Zitvogel L. Cancer despite immunosurveillance: immunoselection and immunosubversion. Nat Rev Immunol. 2006;6:715–27.CrossRefPubMedGoogle Scholar
  4. 4.
    Osusky KL, Hallahan DE, Fu A, et al. The receptor tyrosine kinase inhibitor SU11248 impedes endothelial cell migration, tubule formation, and blood vessel formation in vivo, but has little effect on existing tumor vessels. Angiogenesis. 2004;7:225–33.CrossRefPubMedGoogle Scholar
  5. 5.
    Nemeth JA, Cher ML, Zhou Z, et al. Inhibition of αvβ3 integrin reduces angiogenesis, bone turnover, and tumor cell proliferation in experimental prostate cancer bone metastases. Clin Exp Metastasis. 2003;20:413–20.CrossRefPubMedGoogle Scholar
  6. 6.
    Gotay CC. The experience of cancer during early and advanced stages: the views of patients and their mates. Soc Sci Med. 1984;18:605–13.CrossRefPubMedGoogle Scholar
  7. 7.
    Ostrand-Rosenberg S. Immune surveillance: a balance between pro- and anti-tumor immunity. Curr Opin Genet Dev. 2008;18:11–8.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Gabrilovich DI. Coordinated regulation of myeloid cells by tumours. Nat Rev Immunol. 2012;12:253–68.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Kapp A, McKenna KC. Accumulation of immunosuppressive CD11b+ myeloid cells correlates with the failure to prevent tumor growth in the anterior chamber of the Eye1. J Immunol. 2006;3:1599–608.Google Scholar
  10. 10.
    Mantovani A, Cassatella MA, Costantini C, et al. Neutrophils in the activation and regulation of innate and adaptive immunity. Nat Rev Immunol. 2011;11:519–31.CrossRefPubMedGoogle Scholar
  11. 11.
    Fridlender ZG, Sun J, Kim S, et al. Polarization of tumor-associated neutrophil phenotype by TGF-β: “N1” versus “N2” TAN. Cancer Cell. 2009;16:183–94.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Levy O. Innate immunity of the newborn: basic mechanisms and clinical correlates. Nat Rev Immunol. 2007;5:379–90.CrossRefGoogle Scholar
  13. 13.
    Elifio-Esposito S, Tomazeli L, Schwartz C, et al. Human neutrophil migration and activation by BJcuL, a galactose binding lectin purified from Bothrops jararacussu venom. BMC Immunol. 2011;12:10.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Shojaei F, Singh M, Thompson JD, et al. Role of Bv8 in neutrophil-dependent angiogenesis in a transgenic model of cancer progression. Proc Natl Acad Sci. 2008;105:2640–5.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Tazawa H, Okada F, Kobayashi T, et al. Infiltration of neutrophils is required for acquisition of metastatic phenotype of benign murine fibrosarcoma cells: implication of inflammation-associated carcinogenesis and tumor progression. Am J Pathol. 2003;163:2221–32.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Schmielau J, Finn OJ. Activated granulocytes and granulocyte-derived hydrogen peroxide are the underlying mechanism of suppression of T-cell function in advanced cancer patients. Cancer Res. 2001;61:4756–60.PubMedGoogle Scholar
  17. 17.
    Demers M, Wagner DD. Neutrophil extracellular traps: a new link to cancer-associated thrombosis and potential implications for tumor progression. Oncoimmunology. 2013;2:152–4.CrossRefGoogle Scholar
  18. 18.
    Sceneay J, Chow MT, Chen A, et al. Primary tumor hypoxia recruits CD11b+/Ly6Cmed/Ly6G+ immune suppressor cells and compromises NK cell cytotoxicity in the premetastatic niche. Cancer Res. 2012;72:3906–11.CrossRefPubMedGoogle Scholar
  19. 19.
    Croci DO, Salatino M. Tumor immune escape mechanisms that operate during metastasis. Curr Pharm Biotechnol. 2011;12:1923–36.CrossRefPubMedGoogle Scholar
  20. 20.
    Braun DP, Harris JE. Relationship of leukocyte numbers, immunoregulatory cell function, and phytohemagglutinin responsiveness in cancer patients. J Natl Cancer Inst. 1981;67:809–14.PubMedGoogle Scholar
  21. 21.
    Huang M, Li J, Zhao G, et al. Immunophenotype of myeloid granulocytes: a pilot study for distinguishing myelodysplastic syndrome and aplastic anemia by flow cytometry. Int J Lab Hematol. 2010;32:275–81.CrossRefPubMedGoogle Scholar
  22. 22.
    Fridlender ZG, Sun J, Mishalian I, et al. Transcriptomic analysis comparing tumor-associated neutrophils with granulocytic myeloid-derived suppressor cells and normal neutrophils. PLoS One. 2012;7:812–9.CrossRefGoogle Scholar
  23. 23.
    Hong B, Zu Y. Detecting circulating tumor cells: current challenges and new trends. Theranostics. 2013;3:377–94.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Zitvogel L, Galluzzi L, Smyth MJ, et al. Mechanism of action of conventional and targeted anticancer therapies: reinstating immunosurveillance. Immunity. 2013;39:74–88.CrossRefPubMedGoogle Scholar
  25. 25.
    Granot Z, Henke E, Comen EA, et al. Tumor entrained neutrophils inhibit seeding in the premetastatic lung. Cancer Cell. 2011;20:300–14.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Oliveira AG, Guabiraba R, Teixeira MM, Menezes G. Tumor-associated neutrophils. Trends Stem Cell Prolif Cancer Res 2013;479–501.Google Scholar
  27. 27.
    Rodriguez PC, Hernandez CP, Morrow K, et al. L-arginine deprivation regulates cyclin D3 mRNA stability in human T cells by controlling HuR expression. J Immunol. 2010;185:5198–204.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Smyth MJ, Godfrey DI, Trapani JA. A fresh look at tumor immunosurveillance and immunotherapy. Nat Immunol. 2001;2:293–9.CrossRefPubMedGoogle Scholar
  29. 29.
    Gül N, Babes L, Siegmund K, et al. Macrophages eliminate circulating tumor cells after monoclonal antibody therapy. J Clin Investig. 2014;124:812–23.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Katarina R, Jan HN. CD69 is the crucial regulator of intestinal inflammation: a new target molecule for IBD treatment? J Immunol Res. 2015;2015:1–12.Google Scholar
  31. 31.
    Zhang J, Pan K, Wang W, et al. The prognostic value of tumor-infiltrating neutrophils in gastric adenocarcinoma after resection. PLoS One. 2012;7:e33655.CrossRefGoogle Scholar
  32. 32.
    Maria RG, Cecilia G, Sébastien J, et al. Tumor associated macrophages and neutrophils in tumor progression. J Cell Physiol. 2013;228:1404–12.CrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Institute of Genetics and CytologyNortheast Normal UniversityChangchunChina
  2. 2.Gynecology and Obstetrics DepartmentChina-Japan Union Hospital of Jilin UniversityChangchunChina

Personalised recommendations