Tumor Biology

, Volume 35, Issue 3, pp 2175–2186 | Cite as

HMGB1 combining with tumor-associated macrophages enhanced lymphangiogenesis in human epithelial ovarian cancer

  • Wenqi Zhang
  • Jing Tian
  • Quan Hao
Research Article


Within tumor microenvironment, high-mobility group box protein 1 (HMGB1) and tumor-associated macrophages (TAMs) are able to influence ovarian cancer development and progression via facilitating tumor lymphatic metastasis. However, little is known about the association between HMGB1 and TAMs on lymphangiogenesis in epithelial ovarian cancer (EOC). To investigate the effect of HMGB1 and TAMs on lymphangiogenesis in EOC, immunohistochemistry was performed to determine the expressions of HMGB1, TAMs, and lymphatic vessel density (LVD) in a total of 108 ovarian tissue specimens. Then, the relationships between HMGB1 or TAMs and LVD were assessed by correlation test. In our in vitro study, TAMs were isolated from ascites of EOC patients. Effects of HMGB1, TAMs, and HMGB1 combining with TAMs on lymphatic endothelial cell (LEC) proliferation, migration, and the capillary-like tube formation were measured. Results showed that the expression of HMGB1 and the number of TAMs infiltration were overexpressed in malignant ovarian tumors compared with that in normal ovarian and were closely associated with lymph node metastasis. Positive correlations existed between HMGB1 expression or TAMs count and LVD determination. In an in vitro study, data demonstrated that either HMGB1 or TAMs could facilitate lymphangiogenesis by inducing LEC proliferation, migration, and capillary-like tube formation. Meanwhile, HMGB1 combining with TAMs may augment the pro-lymphangiogenic property. Our data suggest that either HMGB1 or TAMs could facilitate lymphangiogenesis, while HMGB1 coculture with TAMs may strengthen the pro-lymphangiogenic potential, which may serve as a therapeutic target for ovarian cancer.


High-mobility group box 1 protein Tumor-associated macrophages Ovarian cancer Lymphangiogenesis 



This work was supported by a grant-in-aid for scientific research (12JCYBJC17000) from the Tianjin Municipal Science and Technology Commission.

Conflicts of interest



  1. 1.
    Siegel R, Naishadham D, Jemal A. Cancer statistics, 2012. CA Cancer J Clin. 2012;62:10–29.PubMedCrossRefGoogle Scholar
  2. 2.
    Karpanen T, Alitalo K. Lymphatic vessels as targets of tumor therapy? J Exp Med. 2001;194:F37–42.PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Park JS, Gamboni-Robertson F, He Q, Svetkauskaite D, Kim JY, Strassheim D, et al. High mobility group box 1 protein interacts with multiple Toll-like receptors. Am J Physiol Cell Physiol. 2006;290:C917–24.PubMedCrossRefGoogle Scholar
  4. 4.
    Park JS, Svetkauskaite D, He Q, Kim JY, Strassheim D, Ishizaka A, et al. Involvement of toll-like receptors 2 and 4 in cellular activation by high mobility group box 1 protein. J Biol Chem. 2004;279:7370–7.PubMedCrossRefGoogle Scholar
  5. 5.
    Yu M, Wang H, Ding A, Golenbock DT, Latz E, Czura CJ, et al. HMGB1 signals through toll-like receptor (TLR) 4 and TLR2. Shock. 2006;26:174–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Andersson U, Wang H, Palmblad K, Aveberger AC, Bloom O, Erlandsson-Harris H, et al. High mobility group 1 protein (HMG-1) stimulates proinflammatory cytokine synthesis in human monocytes. J Exp Med. 2000;192:565–70.PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Ishiguro H, Nakaigawa N, Miyoshi Y, Fujinami K, Kubota Y, Uemura H. Receptor for advanced glycation end products (RAGE) and its ligand, amphoterin are overexpressed and associated with prostate cancer development. Prostate. 2005;64:92–100.PubMedCrossRefGoogle Scholar
  8. 8.
    Evans A, Lennard TW, Davies BR. High-mobility group protein 1(Y): metastasis-associated or metastasis-inducing? J Surg Oncol. 2004;88:86–99.PubMedCrossRefGoogle Scholar
  9. 9.
    Ellerman JE, Brown CK, de Vera M, Zeh HJ, Billiar T, Rubartelli A, et al. Masquerader: high mobility group box-1 and cancer. Clin Cancer Res. 2007;13:2836–48.PubMedCrossRefGoogle Scholar
  10. 10.
    Tang D, Kang R, Zeh III HJ, Lotze MT. High-mobility group box 1 and cancer. Biochim Biophys Acta. 2010;1799:131–40.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Hao Q, Du XQ, Fu X, Tian J. Expression and clinical significance of HMGB1 and RAGE in cervical squamous cell carcinoma. Zhonghua Zhong Liu Za Zhi [Chin J Oncol]. 2008;30:292–5.Google Scholar
  12. 12.
    Hagemann T, Wilson J, Burke F, Kulbe H, Li NF, Pluddemann A, et al. Ovarian cancer cells polarize macrophages toward a tumor-associated phenotype. J Immunol. 2006;176:5023–32.PubMedCrossRefGoogle Scholar
  13. 13.
    Duluc D, Delneste Y, Tan F, Moles MP, Grimaud L, Lenoir J, et al. Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells. Blood. 2007;110:4319–30.PubMedCrossRefGoogle Scholar
  14. 14.
    Dave SS, Wright G, Tan B, Rosenwald A, Gascoyne RD, Chan WC, et al. Prediction of survival in follicular lymphoma based on molecular features of tumor-infiltrating immune cells. N Engl J Med. 2004;351:2159–69.PubMedCrossRefGoogle Scholar
  15. 15.
    Farinha P, Masoudi H, Skinnider BF, Shumansky K, Spinelli JJ, Gill K, et al. Analysis of multiple biomarkers shows that lymphoma-associated macrophage (LAM) content is an independent predictor of survival in follicular lymphoma (FL). Blood. 2005;106:2169–74.PubMedCrossRefGoogle Scholar
  16. 16.
    Alvaro T, Lejeune M, Salvado MT, Lopez C, Jaen J, Bosch R, et al. Immunohistochemical patterns of reactive microenvironment are associated with clinicobiologic behavior in follicular lymphoma patients. J Clin Oncol. 2006;24:5350–7.PubMedCrossRefGoogle Scholar
  17. 17.
    Kelley T, Beck R, Absi A, Jin T, Pohlman B, Hsi E. Biologic predictors in follicular lymphoma: importance of markers of immune response. Leuk lymphoma. 2007;48:2403–11.PubMedCrossRefGoogle Scholar
  18. 18.
    Zhang QW, Liu L, Gong CY, Shi HS, Zeng YH, Wang XZ, et al. Prognostic significance of tumor-associated macrophages in solid tumor: a meta-analysis of the literature. PloS ONE. 2012;7:e50946.PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Schoppmann SF, Fenzl A, Nagy K, Unger S, Bayer G, Geleff S, et al. VEGF-C expressing tumor-associated macrophages in lymph node positive breast cancer: impact on lymphangiogenesis and survival. Surgery. 2006;139:839–46.PubMedCrossRefGoogle Scholar
  20. 20.
    Gonda TA, Tu S, Wang TC. Chronic inflammation, the tumor microenvironment and carcinogenesis. Cell Cycle. 2009;8:2005–13.PubMedCrossRefGoogle Scholar
  21. 21.
    Mbeunkui F, Johann Jr DJ. Cancer and the tumor microenvironment: a review of an essential relationship. Cancer Chemother Pharmacol. 2009;63:571–82.PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Denholm EM, Wolber FM, Phan SH. Secretion of monocyte chemotactic activity by alveolar macrophages. Am J Pathol. 1989;135:571–80.PubMedCentralPubMedGoogle Scholar
  23. 23.
    Yang S, Cheng H, Cai J, Cai L, Zhang J, Wang Z. PlGF expression in pre-invasive and invasive lesions of uterine cervix is associated with angiogenesis and lymphangiogenesis. APMIS. 2009;117:831–8.PubMedCrossRefGoogle Scholar
  24. 24.
    Mu H, Ohashi R, Yan S, Chai H, Yang H, Lin P, et al. Adipokine resistin promotes in vitro angiogenesis of human endothelial cells. Cardiovasc Res. 2006;70:146–57.PubMedCrossRefGoogle Scholar
  25. 25.
    Gao J, Knutsen A, Arbman G, Carstensen J, Franlund B, Sun XF. Clinical and biological significance of angiogenesis and lymphangiogenesis in colorectal cancer. Dig Liver Dis. 2009;41:116–22.PubMedCrossRefGoogle Scholar
  26. 26.
    Miyata Y, Kanda S, Ohba K, Nomata K, Hayashida Y, Eguchi J, et al. Lymphangiogenesis and angiogenesis in bladder cancer: prognostic implications and regulation by vascular endothelial growth factors-A, -C, and -D. Clin Cancer Res. 2006;12:800–6.PubMedCrossRefGoogle Scholar
  27. 27.
    Tammela T, Alitalo K. Lymphangiogenesis: molecular mechanisms and future promise. Cell. 2010;140:460–76.PubMedCrossRefGoogle Scholar
  28. 28.
    Schoppmann SF, Birner P, Stockl J, Kalt R, Ullrich R, Caucig C, et al. Tumor-associated macrophages express lymphatic endothelial growth factors and are related to peritumoral lymphangiogenesis. Am J Pathol. 2002;161:947–56.PubMedCentralPubMedCrossRefGoogle Scholar
  29. 29.
    Atkins CD. Re: Influence of the new AJCC breast cancer staging system on sentinel lymph node positivity and false-negative rates. J Natl Cancer Inst. 2004;96:1639. author reply 1639–1640.PubMedCrossRefGoogle Scholar
  30. 30.
    Chen J, Xi B, Zhao Y, Yu Y, Zhang J, Wang C. High-mobility group protein B1 (HMGB1) is a novel biomarker for human ovarian cancer. Gynecol Oncol. 2012;126:109–17.PubMedCrossRefGoogle Scholar
  31. 31.
    Qiu Y, Chen Y, Fu X, Zhang L, Tian J, Hao Q. HMGB1 promotes lymphangiogenesis of human lymphatic endothelial cells in vitro. Med Oncol. 2012;29:358–63.PubMedCrossRefGoogle Scholar
  32. 32.
    Grivennikov SI, Greten FR, Karin M. Immunity, inflammation, and cancer. Cell. 2010;140:883–99.PubMedCentralPubMedCrossRefGoogle Scholar
  33. 33.
    Ungefroren H, Sebens S, Seidl D, Lehnert H, Hass R. Interaction of tumor cells with the microenvironment. Cell Commun Signal CCS. 2011;9:18.CrossRefGoogle Scholar
  34. 34.
    Pardo M, Garcia A, Thomas B, Pineiro A, Akoulitchev A, Dwek RA, et al. The characterization of the invasion phenotype of uveal melanoma tumour cells shows the presence of MUC18 and HMG-1 metastasis markers and leads to the identification of DJ-1 as a potential serum biomarker. Int J Cancer. 2006;119:1014–22.PubMedCrossRefGoogle Scholar
  35. 35.
    Volp K, Brezniceanu ML, Bosser S, Brabletz T, Kirchner T, Gottel D, et al. Increased expression of high mobility group box 1 (HMGB1) is associated with an elevated level of the antiapoptotic c-IAP2 protein in human colon carcinomas. Gut. 2006;55:234–42.PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Akaike H, Kono K, Sugai H, Takahashi A, Mimura K, Kawaguchi Y, et al. Expression of high mobility group box chromosomal protein-1 (HMGB-1) in gastric cancer. Anticancer Res. 2007;27:449–57.PubMedGoogle Scholar
  37. 37.
    Biswas SK, Gangi L, Paul S, Schioppa T, Saccani A, Sironi M, et al. A distinct and unique transcriptional program expressed by tumor-associated macrophages (defective NF-kappaB and enhanced IRF-3/STAT1 activation). Blood. 2006;107:2112–22.PubMedCrossRefGoogle Scholar
  38. 38.
    Cao R, Bjorndahl MA, Religa P, Clasper S, Garvin S, Galter D, et al. PDGF-BB induces intratumoral lymphangiogenesis and promotes lymphatic metastasis. Cancer Cell. 2004;6:333–45.PubMedCrossRefGoogle Scholar
  39. 39.
    Fritz-Six KL, Dunworth WP, Li M, Caron KM. Adrenomedullin signaling is necessary for murine lymphatic vascular development. J Clin Investig. 2008;118:40–50.PubMedCentralPubMedCrossRefGoogle Scholar
  40. 40.
    Kajiya K, Hirakawa S, Ma B, Drinnenberg I, Detmar M. Hepatocyte growth factor promotes lymphatic vessel formation and function. EMBO J. 2005;24:2885–95.PubMedCentralPubMedCrossRefGoogle Scholar
  41. 41.
    Scaffidi P, Misteli T, Bianchi ME. Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature. 2002;418:191–5.PubMedCrossRefGoogle Scholar
  42. 42.
    Gardella S, Andrei C, Ferrera D, Lotti LV, Torrisi MR, Bianchi ME, et al. The nuclear protein HMGB1 is secreted by monocytes via a non-classical, vesicle-mediated secretory pathway. EMBO Rep. 2002;3:995–1001.PubMedCentralPubMedCrossRefGoogle Scholar
  43. 43.
    Venneri MA, De Palma M, Ponzoni M, Pucci F, Scielzo C, Zonari E, et al. Identification of proangiogenic TIE2-expressing monocytes (TEMs) in human peripheral blood and cancer. Blood. 2007;109:5276–85.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2013

Authors and Affiliations

  1. 1.Department of Gynecologic OncologyTianjin Medical University Cancer Institute and Hospital, National Clinical Research Centre of CancerTianjinChina
  2. 2.Key Laboratory of Cancer Prevention and TherapyTianjinChina

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