Tumor Biology

, Volume 36, Issue 2, pp 1163–1171 | Cite as

Higher circulating levels of chemokine CCL22 in patients with breast cancer: evaluation of the influences of tumor stage and chemokine gene polymorphism

  • A. Jafarzadeh
  • H. Fooladseresht
  • K. Minaee
  • M. R. Bazrafshani
  • A. Khosravimashizi
  • M. Nemati
  • M. Mohammadizadeh
  • M. M. Mohammadi
  • A. Ghaderi
Research Article


The receptor for CCL22 is named CCR4 that preferentially is expressed on the regulatory T cells (Treg), and accordingly, CCL22 acts as a chemoattractant for the intratumoral Treg migration. The aim of this study was to evaluate the serum CCL22 levels and a single nucleotide polymorphism (SNP) in chemokine gene, [2030 G/C (rs223818)], in patients with breast cancer. Blood samples were collected from 100 women with breast cancer before receiving chemotherapy, radiotherapy, or immunotherapy and 100 age-matched healthy women as a control group. The serum CCL22 levels were measured by ELISA. The DNA extracted and the SNP rs223818 determined by amplification refractory mutation system–polymerase chain reaction (ARMS–PCR) technique. The mean serum CCL22 levels in patients with breast cancer (2398.5 ± 123 Pg/mL) was significantly higher in comparison to healthy control group (974.2 ± 39.9 Pg/mL; P < 0.001). According to the tumor stages, the mean serum levels of CCL22 were 999.8 ± 85.0 Pg/mL in stage I, 1718.8 ± 82.3 Pg/mL in stage II, 2846.8 ± 118.0 Pg/mL in stage III, and 3954.5 ± 245.2 Pg/mL in stage IV. There was significant difference between tumor stages regarding the serum CCL22 levels (P < 0.001). In patients with breast cancer, the frequencies of CC genotype (63 %) and C allele (79 %) at rs223818 were significantly higher as compared to healthy controls (31 and 52 %, respectively; P < 0.001). In both patients and control groups, the mean serum levels of CCL22 in subjects with CC genotype or C allele at rs223818 were also significantly higher as compared to subjects with GG genotype or G allele (P < 0.001). Higher serum CCL22 levels were observed in patients with breast cancer that is increased with advanced stages. These findings represent that the CCL22 may contribute in tumor development. The CC genotype and C allele at rs223818 were more frequent in breast cancer patients. The serum CCL22 levels were affected by genetic variations at SNP rs223818. Accordingly, SNP rs223818 may play a role in the susceptibility to breast cancer.


Chemokine CCL22 Breast cancer Gene polymorphism rs223818 


Conflicts of interest



  1. 1.
    Nelson HD, Zakher B, Cantor A, Fu R, Griffin J, O’Meara ES, et al. Risk factors for breast cancer for women aged 40 to 49 years: a systematic review and meta-analysis. Ann Intern Med. 2012;156:635–48.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin. 2013;63:11–30.CrossRefPubMedGoogle Scholar
  3. 3.
    Park NJ, Kang DH. Breast cancer risk and immune responses in healthy women. Oncol Nurs Forum. 2006;33:1151–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Kees T, Egeblad M. Innate immune cells in breast cancer—from villains to heroes? J Mammary Gland Biol Neoplasia. 2011;16:189–203.CrossRefPubMedGoogle Scholar
  5. 5.
    Gruber I, Landenberger N, Staebler A, Hahn M, Wallwiener D, Fehm T. Relationship between circulating tumor cells and peripheral T-cells in patients with primary breast cancer. Anticancer Res. 2013;33:2233–8.PubMedGoogle Scholar
  6. 6.
    Zhu J, Paul WE. Heterogeneity and plasticity of T helper cells. Cell Res. 2010;20:4–12.CrossRefPubMedGoogle Scholar
  7. 7.
    Faghih Z, Erfani N, Haghshenas MR, Safaei A, Talei AR, Ghaderi A. Immune profiles of CD4+ lymphocyte subsets in breast cancer tumor draining lymph nodes. Immunol Lett. 2014;158:57–65.CrossRefPubMedGoogle Scholar
  8. 8.
    Kennedy R, Celis E. Multiple roles for CD4+ T cells in anti-tumor immune responses. Immunol Rev. 2008;222:129–44.CrossRefPubMedGoogle Scholar
  9. 9.
    Lee HJ, Song IC, Yun HJ, Jo DY, Kim S. CXC chemokines and chemokine receptors in gastric cancer: from basic findings towards therapeutic targeting. World J Gastroenterol. 2014;20:1681–93.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Li J-Y, Ou Z-L, Yu S-J, Gu X-L, Yang C, Chen A-X, et al. The chemokine receptor CCR4 promotes tumor growth and lung metastasis in breast cancer. Breast Cancer Res Treat. 2012;131:837–48.CrossRefPubMedGoogle Scholar
  11. 11.
    Zhu Q, Han X, Peng J, Qin H, Wang Y. The role of CXC chemokines and their receptors in the progression and treatment of tumors. J Mol Histol. 2012;43:699–713.CrossRefPubMedGoogle Scholar
  12. 12.
    Vandercappellen J, Van Damme J, Struyf S. The role of CXC chemokines and their receptors in cancer. Cancer Lett. 2008;267:226–44.CrossRefPubMedGoogle Scholar
  13. 13.
    Viola A, Sarukhan A, Bronte V, Molon B. The pros and cons of chemokines in tumor immunology. Trends Immunol. 2012;33:496–504.CrossRefPubMedGoogle Scholar
  14. 14.
    Franciszkiewicz K, Boissonnas A, Boutet M, Combadiere C, Mami-Chouaib F. Role of chemokines and chemokine receptors in shaping the effector phase of the antitumor immune response. Cancer Res. 2012;72:6325–32.CrossRefPubMedGoogle Scholar
  15. 15.
    Yamashita U, Kuroda E. Regulation of macrophage-derived chemokine (MDC, CCL22) production. Crit Rev Immunol. 2002;22:105–14.CrossRefPubMedGoogle Scholar
  16. 16.
    Wang G, Yu D, Tan W, Zhao D, Wu C, Lin D. Genetic polymorphism in chemokine CCL22 and susceptibility to Helicobacter pylori infection‐related gastric carcinoma. Cancer. 2009;115:2430–7.CrossRefPubMedGoogle Scholar
  17. 17.
    Nishikawa H, Sakaguchi S. Regulatory T cells in tumor immunity. Int J Cancer. 2010;127:759–67.PubMedGoogle Scholar
  18. 18.
    Wagsater D, Dienus O, Lofgren S, Hugander A, Dimberg J. Quantification of the chemokines CCL17 and CCL22 in human colorectal adenocarcinomas. Mol Med Rep. 2008;1:211–7.PubMedGoogle Scholar
  19. 19.
    Nakanishi T, Imaizumi K, Hasegawa Y, Kawabe T, Hashimoto N, Okamoto M, et al. Expression of macrophage-derived chemokine (MDC)/CCL22 in human lung cancer. Cancer Immunol Immunother. 2006;55:1320–9.CrossRefPubMedGoogle Scholar
  20. 20.
    Li YQ, Liu FF, Zhang XM, Guo XJ, Ren MJ, Fu L. Tumor secretion of CCL22 activates intratumoral Treg infiltration and is independent prognostic predictor of breast cancer. PLoS One. 2013;8:e76379.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Niens M, Visser L, Nolte IM, Van Der Steege G, Diepstra A, Cordano P, et al. Serum chemokine levels in Hodgkin lymphoma patients: highly increased levels of CCL17 and CCL22. Br J Haematol. 2008;140:527–36.CrossRefPubMedGoogle Scholar
  22. 22.
    Tsujikawa T, Yaguchi T, Ohmura G, Ohta S, Kobayashi A, Kawamura N, et al. Autocrine and paracrine loops between cancer cells and macrophages promote lymph node metastasis via CCR4/CCL22 in head and neck squamous cell carcinoma. Int J Cancer. 2013;132:2755–66.CrossRefPubMedGoogle Scholar
  23. 23.
    Sugimoto M, Yamaoka Y, Furuta T. Influence of interleukin polymorphisms on development of gastric cancer and peptic ulcer. World J Gastroenterol. 2010;16:1188–200.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    You Y, Deng J, Zheng J, Hu M, Li N, Wu H, et al. IL-21 gene polymorphism is associated with the prognosis of breast cancer in Chinese populations. Breast Cancer Res Treat. 2013;137:893–901.CrossRefPubMedGoogle Scholar
  25. 25.
    Back LK, Farias TD, da Cunha PA, Muniz YC, Ribeiro MC, Fernandes BL, et al. Functional polymorphisms of interleukin-18 gene and risk of breast cancer in a Brazilian population. Tissue Antigens. 2014;2014:12367.Google Scholar
  26. 26.
    Karakus N, Kara N, Ulusoy AN, Ozaslan C, Bek Y. Tumor necrosis factor alpha and beta and interferon gamma gene polymorphisms in Turkish breast cancer patients. DNA Cell Biol. 2011;30:371–7.CrossRefPubMedGoogle Scholar
  27. 27.
    Ma XY, Jin Y, Sun HM, Yu L, Bai J, Chen F, et al. CXCL12 G801A polymorphism contributes to cancer susceptibility: a meta-analysis. Cell Mol Biol (Noisy-le-grand). 2012;58(Suppl):OL1702–1708.Google Scholar
  28. 28.
    Guergnon J, Combadiere C. Role of chemokines polymorphisms in diseases. Immunol Lett. 2012;145:15–22.CrossRefPubMedGoogle Scholar
  29. 29.
    Tahmasebi Z, Akbarian M, Mirkazemi S, Shahlaee A, Alizadeh Z, Amirzargar AA, et al. Interleukin-1 gene cluster and IL-1 receptor polymorphisms in Iranian patients with systemic lupus erythematosus. Rheumatol Int. 2013;33:2591–6.CrossRefPubMedGoogle Scholar
  30. 30.
    Wang G, Yu D, Tan W, Zhao D, Wu C, Lin D. Genetic polymorphism in chemokine CCL22 and susceptibility to Helicobacter pylori infection-related gastric carcinoma. Cancer. 2009;115:2430–7.CrossRefPubMedGoogle Scholar
  31. 31.
    Hirota T, Saeki H, Tomita K, Tanaka S, Ebe K, Sakashita M, et al. Variants of CC motif chemokine 22 (CCL22) are associated with susceptibility to atopic dermatitis: case–control studies. PLoS One. 2011;6.Google Scholar
  32. 32.
    Greene F, Page D, Fleming I, Fritz A, Balch C, Haller D, et al. R C: AJCC cancer staging man. NY: Springer; 2002.CrossRefGoogle Scholar
  33. 33.
    Miller S, Dykes D, Polesky H. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988;16:1215.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Perrey C, Turner SJ, Pravica V, Howell WM, Hutchinson IV. ARMS-PCR methodologies to determine IL-10, TNF-α, TNF-β and TGF-β1 gene polymorphisms. Transpl Immunol. 1999;7:127–8.CrossRefPubMedGoogle Scholar
  35. 35.
    Hallett MA, Venmar KT, Fingleton B. Cytokine stimulation of epithelial cancer cells: the similar and divergent functions of IL-4 and IL-13. Cancer Res. 2012;72:6338–43.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Shimauchi T, Imai S, Hino R, Tokura Y. Production of thymus and activation-regulated chemokine and macrophage-derived chemokine by CCR4+ adult T-cell leukemia cells. Clin Cancer Res. 2005;11:2427–35.CrossRefPubMedGoogle Scholar
  37. 37.
    Takegawa S, Jin Z, Nakayama T, Oyama T, Hieshima K, Nagakubo D, et al. Expression of CCL17 and CCL22 by latent membrane protein 1-positive tumor cells in age-related Epstein–Barr virus-associated B-cell lymphoproliferative disorder. Cancer Sci. 2008;99:296–302.CrossRefPubMedGoogle Scholar
  38. 38.
    Faget J, Biota C, Bachelot T, Gobert M, Treilleux I, Goutagny N, et al. Early detection of tumor cells by innate immune cells leads to Treg recruitment through CCL22 production by tumor cells. Cancer Res. 2011;71:6143–52.CrossRefPubMedGoogle Scholar
  39. 39.
    Menetrier-Caux C, Faget J, Biota C, Gobert M, Blay JY, Caux C. Innate immune recognition of breast tumor cells mediates CCL22 secretion favoring Treg recruitment within tumor environment. Oncoimmunology. 2012;1:759–61.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Mailloux AW, Young MR. NK-dependent increases in CCL22 secretion selectively recruits regulatory T cells to the tumor microenvironment. J Immunol. 2009;182:2753–65.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Fialova A, Partlova S, Sojka L, Hromadkova H, Brtnicky T, Fucikova J, et al. Dynamics of T-cell infiltration during the course of ovarian cancer: the gradual shift from a Th17 effector cell response to a predominant infiltration by regulatory T-cells. Int J Cancer. 2013;132:1070–9.CrossRefPubMedGoogle Scholar
  42. 42.
    Miller AM, Lundberg K, Ozenci V, Banham AH, Hellstrom M, Egevad L, et al. CD4 + CD25high T cells are enriched in the tumor and peripheral blood of prostate cancer patients. J Immunol. 2006;177:7398–405.CrossRefPubMedGoogle Scholar
  43. 43.
    Mizukami Y, Kono K, Kawaguchi Y, Akaike H, Kamimura K, Sugai H, et al. CCL17 and CCL22 chemokines within tumor microenvironment are related to accumulation of Foxp3+ regulatory T cells in gastric cancer. Int J Cancer. 2008;122:2286–93.CrossRefPubMedGoogle Scholar
  44. 44.
    Maruyama T, Kono K, Izawa S, Mizukami Y, Kawaguchi Y, Mimura K, et al. CCL17 and CCL22 chemokines within tumor microenvironment are related to infiltration of regulatory T cells in esophageal squamous cell carcinoma. Dis Esophagus. 2010;23:422–9.PubMedGoogle Scholar
  45. 45.
    Benevides L, Cardoso CR, Tiezzi DG, Marana HR, Andrade JM, Silva JS. Enrichment of regulatory T cells in invasive breast tumor correlates with the upregulation of IL-17A expression and invasiveness of the tumor. Eur J Immunol. 2013;43:1518–28.CrossRefPubMedGoogle Scholar
  46. 46.
    Whiteside TL. Regulatory T cell subsets in human cancer: are they regulating for or against tumor progression? Cancer Immunol Immunother. 2013;63:67–72.CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Beaty SR, Rose Jr CE, Sung SS. Diverse and potent chemokine production by lung CD11bhigh dendritic cells in homeostasis and in allergic lung inflammation. J Immunol. 2007;178:1882–95.CrossRefPubMedGoogle Scholar
  48. 48.
    Wang ZK, Yang B, Liu H, Hu Y, Yang JL, Wu LL, et al. Regulatory T cells increase in breast cancer and in stage IV breast cancer. Cancer Immunol Immunother. 2012;61:911–6.CrossRefPubMedGoogle Scholar
  49. 49.
    Zhao Y, Wu K, Cai K, Zhai R, Tao K, Wang G, et al. Increased numbers of gastric-infiltrating mast cells and regulatory T cells are associated with tumor stage in gastric adenocarcinoma patients. Oncol Lett. 2012;4:755–8.PubMedPubMedCentralGoogle Scholar
  50. 50.
    Bacic D, Uravic M, Bacic R, Sutic I, Petrosic N. Augmentation of regulatory T cells (CD4 + CD25 + Foxp3+) correlates with tumor stage in patients with colorectal cancer. Coll Antropol. 2011;35 Suppl 2:65–8.PubMedGoogle Scholar
  51. 51.
    Antonelli A, Ferrari SM, Giuggioli D, Ferrannini E, Ferri C, Fallahi P. Chemokine (C-X-C motif) ligand (CXCL)10 in autoimmune diseases. Autoimmun Rev. 2014;13:272–80.CrossRefPubMedGoogle Scholar
  52. 52.
    Liu M, Guo S, Hibbert JM, Jain V, Singh N, Wilson NO, et al. CXCL10/IP-10 in infectious diseases pathogenesis and potential therapeutic implications. Cytokine Growth Factor Rev. 2011;22:121–30.PubMedPubMedCentralGoogle Scholar
  53. 53.
    Mulligan AM, Raitman I, Feeley L, Pinnaduwage D, Nguyen LT, O’Malley FP, et al. Tumoral lymphocytic infiltration and expression of the chemokine CXCL10 in breast cancers from the Ontario familial breast cancer registry. Clin Cancer Res. 2013;19:336–46.CrossRefPubMedGoogle Scholar
  54. 54.
    Haabeth OA, Lorvik KB, Hammarstrom C, Donaldson IM, Haraldsen G, Bogen B, et al. Inflammation driven by tumour-specific Th1 cells protects against B-cell cancer. Nat Commun. 2011;2:240.CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Li G, Tian L, Hou JM, Ding ZY, He QM, Feng P, et al. Improved therapeutic effectiveness by combining recombinant CXC chemokine ligand 10 with cisplatin in solid tumors. Clin Cancer Res. 2005;11:4217–24.CrossRefPubMedGoogle Scholar
  56. 56.
    Chew V, Chen J, Lee D, Loh E, Lee J, Lim KH, et al. Chemokine-driven lymphocyte infiltration: an early intratumoural event determining long-term survival in resectable hepatocellular carcinoma. Gut. 2012;61:427–38.CrossRefPubMedGoogle Scholar
  57. 57.
    Mlecnik B, Tosolini M, Charoentong P, Kirilovsky A, Bindea G, Berger A, et al. Biomolecular network reconstruction identifies T-cell homing factors associated with survival in colorectal cancer. Gastroenterology. 2010;138:1429–40.CrossRefPubMedGoogle Scholar
  58. 58.
    Cooper DN. Functional intronic polymorphisms: buried treasure awaiting discovery within our genes. Hum Genom. 2010;4:284–8.CrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • A. Jafarzadeh
    • 1
    • 2
  • H. Fooladseresht
    • 1
  • K. Minaee
    • 1
  • M. R. Bazrafshani
    • 3
  • A. Khosravimashizi
    • 1
  • M. Nemati
    • 1
  • M. Mohammadizadeh
    • 4
  • M. M. Mohammadi
    • 2
  • A. Ghaderi
    • 5
    • 6
  1. 1.Department of Immunology, Medical SchoolKerman University of Medical SciencesKermanIran
  2. 2.Department of Immunology, Medical SchoolRafsanjan University of Medical SciencesRafsanjanIran
  3. 3.Department of Medical Genetic, Medical SchoolKerman University of Medical SciencesKermanIran
  4. 4.Department of Hematology, Medical SchoolKerman University of Medical SciencesKermanIran
  5. 5.Shiraz Institute for Cancer ResearchShiraz University of Medical SciencesShirazIran
  6. 6.Department of Immunology, School of MedicineShiraz University of Medical SciencesShirazIran

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