Skip to main content

Advertisement

SpringerLink
Log in

T-box transcription factor Brachyury in lung cancer cells inhibits macrophage infiltration by suppressing CCL2 and CCL4 chemokines

  • Research Article
  • Published:
Tumor Biology

Abstract

Both intra-tumor macrophage and T-box transcription factor Brachyury (T) have been proved to play important roles in tumor progression and metastasis. However, it is still unknown whether T could regulate the infiltration of macrophages. Here, we report that the Brachyury expression in human lung tumors was inversely correlated with the infiltration of macrophages. Brachyury suppressed the capability of human lung cancer cells to attract macrophages. Using PCR array, we found that Brachyury inhibited expression of several chemokines, including CCL2, CCL4, and CXCL10. Accordingly, knockdown of CCL2 and CCL4 in lung cancer cells suppressed macrophage invasion under coculture conditions. Furthermore, we found that Brachyury expression was inversely correlated with CCL2 and CCL4 expression in human lung tumors. Taken together, our findings shed light on the novel role of Brachyury in regulation of macrophage infiltration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA Cancer J Clin. 2014;64:9–29.

    Article  PubMed  Google Scholar 

  2. Cirkel GA, Gadellaa-van Hooijdonk CG, Koudijs MJ, Willems SM, Voest EE. Tumor heterogeneity and personalized cancer medicine: are we being outnumbered? Future Oncol. 2014;10:417–28.

    Article  PubMed  Google Scholar 

  3. Longo DL. Tumor heterogeneity and personalized medicine. N Engl J Med. 2012;366:956–7.

    Article  CAS  PubMed  Google Scholar 

  4. Wood SL, Pernemalm M, Crosbie PA, Whetton AD. The role of the tumor-microenvironment in lung cancer-metastasis and its relationship to potential therapeutic targets. Cancer Treat Rev. 2014;40:558–66.

    Article  CAS  PubMed  Google Scholar 

  5. Eming SA, Krieg T, Davidson JM. Inflammation in wound repair: molecular and cellular mechanisms. J Invest Dermatol. 2007;127:514–25.

    Article  CAS  PubMed  Google Scholar 

  6. Ovchinnikov DA. Macrophages in the embryo and beyond: much more than just giant phagocytes. Genesis. 2008;46:447–62.

    Article  PubMed  Google Scholar 

  7. Mills CD. M1 and m2 macrophages: oracles of health and disease. Crit Rev Immunol. 2012;32:463–88.

    Article  CAS  PubMed  Google Scholar 

  8. Deodhar AK, Rana RE. Surgical physiology of wound healing: a review. J Postgrad Med. 1997;43:52–6.

    CAS  PubMed  Google Scholar 

  9. Greenhalgh DG. The role of apoptosis in wound healing. Int J Biochem Cell Biol. 1998;30:1019–30.

    Article  CAS  PubMed  Google Scholar 

  10. Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic-Canic M. Growth factors and cytokines in wound healing. Wound Repair Regen. 2008;16:585–601.

    Article  PubMed  Google Scholar 

  11. Papaioannou VE. T-box genes in development: from hydra to humans. Int Rev Cytol. 2001;207:1–70.

    Article  CAS  PubMed  Google Scholar 

  12. Tada M, Smith JC. T-targets: clues to understanding the functions of t-box proteins. Dev Growth Differ. 2001;43:1–11.

    Article  CAS  PubMed  Google Scholar 

  13. Dobrovolskaia-Zavadskaia N. Regarding the spontaneous mortification of the tail of a new-born mouse and the existence of a hereditary characteristic (factor). C R Seances Soc Biol Fil. 1927;97:114–6.

    Google Scholar 

  14. Wilkinson DG, Bhatt S, Herrmann BG. Expression pattern of the mouse t gene and its role in mesoderm formation. Nature. 1990;343:657–9.

    Article  CAS  PubMed  Google Scholar 

  15. Klaus A, Birchmeier W. Wnt signalling and its impact on development and cancer. Nat Rev Cancer. 2008;8:387–98.

    Article  CAS  PubMed  Google Scholar 

  16. 16Li Y, Luo H, Liu T, Zacksenhaus E, Ben-David Y. The ets transcription factor fli-1 in development, cancer and disease. Oncogene. 2014.

  17. Hsu I, Vitkus S, Da J, Yeh S. Role of oestrogen receptors in bladder cancer development. Nat Rev Urol. 2013;10:317–26.

    Article  CAS  PubMed  Google Scholar 

  18. Takebe N, Nguyen D, Yang SX. Targeting notch signaling pathway in cancer: clinical development advances and challenges. Pharmacol Ther. 2014;141:140–9.

    Article  CAS  PubMed  Google Scholar 

  19. Turner N, Grose R. Fibroblast growth factor signalling: from development to cancer. Nat Rev Cancer. 2010;10:116–29.

    Article  CAS  PubMed  Google Scholar 

  20. Cao L, Yu Y, Bilke S, Walker RL, Mayeenuddin LH, Azorsa DO, et al. Genome-wide identification of pax3-fkhr binding sites in rhabdomyosarcoma reveals candidate target genes important for development and cancer. Cancer Res. 2010;70:6497–508.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Thoma C. Prostate cancer: Brachyury—a biomarker for progression and prognosis? Nat Rev Urol. 2014.

  22. Pires MM, Aaronson SA. Brachyury: a new player in promoting breast cancer aggressiveness. J Natl Cancer Inst. 2014;106.

  23. Pinto F, Pertega-Gomes N, Pereira MS, Vizcaino JR, Monteiro P, Henrique RM, et al. T-box transcription factor brachyury is associated with prostate cancer progression and aggressiveness. Clin Cancer Res. 2014.

  24. Palena C, Roselli M, Litzinger MT, Ferroni P, Costarelli L, Spila A, et al. Overexpression of the emt driver brachyury in breast carcinomas: association with poor prognosis. J Natl Cancer Inst. 2014;106.

  25. Kobayashi Y, Sugiura T, Imajyo I, Shimoda M, Ishii K, Akimoto N, et al. Knockdown of the t-box transcription factor brachyury increases sensitivity of adenoid cystic carcinoma cells to chemotherapy and radiation in vitro: Implications for a new therapeutic principle. Int J Oncol. 2014;44:1107–17.

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Roselli M, Fernando RI, Guadagni F, Spila A, Alessandroni J, Palmirotta R, et al. Brachyury, a driver of the epithelial-mesenchymal transition, is overexpressed in human lung tumors: an opportunity for novel interventions against lung cancer. Clin Cancer Res. 2012;18:3868–79.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Park JC, Chae YK, Son CH, Kim MS, Lee J, Ostrow K, et al. Epigenetic silencing of human t (brachyury homologue) gene in non-small-cell lung cancer. Biochem Biophys Res Commun. 2008;365:221–6.

    Article  CAS  PubMed  Google Scholar 

  28. Singer JB, Harbecke R, Kusch T, Reuter R, Lengyel JA. Drosophila brachyenteron regulates gene activity and morphogenesis in the gut. Development. 1996;122:3707–18.

    CAS  PubMed  Google Scholar 

  29. Zheng J, Yang M, Shao J, Miao Y, Han J, Du J. Chemokine receptor cx3cr1 contributes to macrophage survival in tumor metastasis. Mol Cancer. 2013;12:141.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Iida N, Nakamoto Y, Baba T, Nakagawa H, Mizukoshi E, Naito M, et al. Antitumor effect after radiofrequency ablation of murine hepatoma is augmented by an active variant of cc chemokine ligand 3/macrophage inflammatory protein-1alpha. Cancer Res. 2010;70:6556–65.

    Article  CAS  PubMed  Google Scholar 

  31. Mizutani K, Sud S, McGregor NA, Martinovski G, Rice BT, Craig MJ, et al. The chemokine ccl2 increases prostate tumor growth and bone metastasis through macrophage and osteoclast recruitment. Neoplasia. 2009;11:1235–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Herrmann BG, Labeit S, Poustka A, King TR, Lehrach H. Cloning of the t gene required in mesoderm formation in the mouse. Nature. 1990;343:617–22.

    Article  CAS  PubMed  Google Scholar 

  33. Dobrovolskaia-Zavadskaia N. Brachyura, accompanying the bendings and the genetic structure of the tail of the mouse. C R Seances Soc Biol Fil. 1927;97:1583–5.

    Google Scholar 

  34. Yamada A, Koyanagi KO, Watanabe H. In silico and in vivo identification of the intermediate filament vimentin that is downregulated downstream of brachyury during xenopus embryogenesis. Gene. 2012;491:232–6.

    Article  CAS  PubMed  Google Scholar 

  35. Katikala L, Aihara H, Passamaneck YJ, Gazdoiu S, Jose-Edwards DS, Kugler JE, et al. Functional brachyury binding sites establish a temporal read-out of gene expression in the ciona notochord. PLoS Biol. 2013;11:e1001697.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Kilic N, Feldhaus S, Kilic E, Tennstedt P, Wicklein D, Wasielewski R, et al. Brachyury expression predicts poor prognosis at early stages of colorectal cancer. Eur J Cancer. 2011;47:1080–5.

    Article  PubMed  Google Scholar 

  37. Sarkar D, Shields B, Davies ML, Muller J, Wakeman JA. Brachyury confers cancer stem cell characteristics on colorectal cancer cells. Int J Cancer. 2012;130:328–37.

    Article  CAS  PubMed  Google Scholar 

  38. Pillay N, Plagnol V, Tarpey PS, Lobo SB, Presneau N, Szuhai K, et al. A common single-nucleotide variant in t is strongly associated with chordoma. Nat Genet. 2012;44:1185–7.

    Article  CAS  PubMed  Google Scholar 

  39. Kitamura Y, Sasaki H, Kimura T, Miwa T, Takahashi S, Kawase T, et al. Molecular and clinical risk factors for recurrence of skull base chordomas: gain on chromosome 2p, expression of brachyury, and lack of irradiation negatively correlate with patient prognosis. J Neuropathol Exp Neurol. 2013;72:816–23.

    Article  CAS  PubMed  Google Scholar 

  40. Nibu Y, Jose-Edwards DS, Di Gregorio A. From notochord formation to hereditary chordoma: the many roles of brachyury. Biomed Res Int. 2013;2013:826435.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Barresi V, Ieni A, Branca G, Tuccari G. Brachyury: a diagnostic marker for the differential diagnosis of chordoma and hemangioblastoma versus neoplastic histological mimickers. Dis Markers. 2014;2014:514753.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Scheil-Bertram S, Kappler R, von Baer A, Hartwig E, Sarkar M, Serra M, et al. Molecular profiling of chordoma. Int J Oncol. 2014;44:1041–55.

    PubMed  PubMed Central  Google Scholar 

  43. Haro A, Yano T, Kohno M, Yoshida T, Koga T, Okamoto T, et al. Expression of brachyury gene is a significant prognostic factor for primary lung carcinoma. Ann Surg Oncol. 2013;20 Suppl 3:S509–516.

    Article  PubMed  Google Scholar 

  44. Quatromoni JG, Eruslanov E. Tumor-associated macrophages: function, phenotype, and link to prognosis in human lung cancer. Am J Transl Res. 2012;4:376–89.

    PubMed  PubMed Central  Google Scholar 

  45. Becker M, Muller CB, De Bastiani MA, Klamt F. The prognostic impact of tumor-associated macrophages and intra-tumoral apoptosis in non-small cell lung cancer. Histol Histopathol. 2014;29:21–31.

    PubMed  Google Scholar 

  46. El-Nikhely N, Larzabal L, Seeger W, Calvo A, Savai R. Tumor-stromal interactions in lung cancer: novel candidate targets for therapeutic intervention. Expert Opin Investig Drugs. 2012;21:1107–22.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study Project is supported by the National Natural Science Foundation of China (Grant No. 81102036)

Conflicts of interest

None

Author information

Author notes

    Authors and Affiliations

    1. Department of Bone Tumor Surgery, Changzheng Hospital, Second Military Medical University, NO. 415 Fengyang Road, Shanghai, 200003, China

      Su Chen, Jian Jiao, Dongjie Jiang, Zongmiao Wan, Leqin Xu, Zhenhua Zhou, Wei Xu & Jianru Xiao

    2. Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences, East China Normal University, Shanghai, 200241, China

      Lei Li & Ke Li

    Authors
    1. Su Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Jian Jiao
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Dongjie Jiang
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Zongmiao Wan
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Lei Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Ke Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Leqin Xu
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Zhenhua Zhou
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Wei Xu
      View author publications

      You can also search for this author in PubMed Google Scholar

    10. Jianru Xiao
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Jianru Xiao.

    Additional information

    Su Chen and Jian Jiao contributed equally to this work.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    Supplementary Table 1

    Patient information of the specimens enrolled in the study. Case numbers were determined upon hospitalization. M stands for male; F stands for female and the ages were as shown. The diagnosis and the location of the tumors are recorded. TNM stage was determined equivalent to American Joint Committee of Cancer-AJCC staging standard. Scoring of the IHC staining was recorded as the average score from three independent pathologists’ analysis. (PDF 15 kb)

    Supplementary Figure 1

    Differential expression level of Brachyury in lung cancer cell lines. A. Semi-quantitative RT-PCR was done to test the expression level in the lung cancer cell lines as indicated. Samples were retrieved after 30 cycles of PCR amplification. 18S ribosome RNA was used as loading control to ensure equal loading of each sample. B. Western Blot result of Brachyury expression in the cell lines shown. GAPDH was blotted as loading control. (PDF 30 kb)

    Supplementary Figure 2

    Generation of H460 cells with stable knockdown of BrachyuryandH1299 cell line with overexpression of Brachyury. A. Semi-quantitative RT-PCR of Brachyury expression in H460 cells transfected with control shRNAs (H460shN) or Brachyury shRNAs (H460shT). 18S was used as loading control. B. Western blot result testing Brachyury expression in H460shN and H460shT cells. GAPDH was used as loading control. C. RT-PCR examination of Brachyury expression in H1299 cells transfected with empty vector (H1299EV) or Brachyury overexpression plasmid (H1299TOE). 18S was used as loading control. D. Western blot result testing Brachyury expression in H1299EV and H1299TOE cells. GAPDH was used as loading control. (PDF 34 kb)

    Supplementary Figure 3

    The gene list of 84 genes that encode chemokines and their receptors (PDF 13 kb)

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Chen, S., Jiao, J., Jiang, D. et al. T-box transcription factor Brachyury in lung cancer cells inhibits macrophage infiltration by suppressing CCL2 and CCL4 chemokines. Tumor Biol. 36, 5881–5890 (2015). https://doi.org/10.1007/s13277-015-3260-2

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s13277-015-3260-2

    Keywords

    Search

    Navigation