Virchows Archiv

, Volume 453, Issue 3, pp 257–265 | Cite as

The role of VEGF-C staining in predicting regional metastasis in melanoma

  • Barbara Boone
  • Willeke Blokx
  • Dirk De Bacquer
  • Jo Lambert
  • Dirk Ruiter
  • Lieve Brochez
Original Article

Abstract

Sentinel lymph node status is the most important prognostic factor in primary melanoma. The number of melanoma-associated lymphatic vessels has been associated with sentinel lymph node status and survival. Vascular endothelial growth factor-C (VEGF-C) is found to promote tumour-associated lymphatic vessel growth. In many human neoplasms, VEGF-C expression in neoplastic cells or tumour-associated macrophages (TAMs) has been linked to lymphatic dissemination of tumour cells. Recent studies have suggested a correlation between VEGF-C expression in primary melanoma and the presence of lymph node metastasis. We performed VEGF-C immunohistochemical staining on melanoma tissues of 113 patients with known sentinel lymph node status. We showed that both high VEGF-C expression in melanoma cells and TAMs are positively associated with the presence of a positive sentinel lymph node. No correlation with Breslow thickness, Clark invasion level or ulceration could be detected. VEGF-C expression in melanoma cells was predictive of a shorter overall and disease-free survival, without being an independent predictor of survival. Our results confirm that VEGF-C expression in primary cutaneous melanoma plays a role in the lymphatic spread of the tumour.

Keywords

Skin Melanoma Lymphangiogenesis Sentinel lymph node VEGF-C TAMs Immunohistochemistry 

References

  1. 1.
    Balch CM, Buzaid AC, Soong SJ, Atkins MB, Cascinelli N, Coit DG, Fleming ID, Gershenwald JE, Houghton A Jr., Kirkwood JM, McMasters KM, Mihm MF, Morton DL, Reintgen DS, Ross MI, Sober A, Thompson JA, Thompson JF (2001) Final version of the American Joint Committee on Cancer staging system for cutaneous melanoma. J Clin Oncol 19:3635–3648PubMedGoogle Scholar
  2. 2.
    Balch CM, Soong SJ, Gershenwald JE, Thompson JF, Reintgen DS, Cascinelli N, Urist M, McMasters KM, Ross MI, Kirkwood JM, Atkins MB, Thompson JA, Coit DG, Byrd D, Desmond R, Zhang Y, Liu PY, Lyman GH, Morabito A (2001) Prognostic factors analysis of 17,600 melanoma patients: validation of the American Joint Committee on Cancer melanoma staging system. J Clin Oncol 19:3622–3634PubMedGoogle Scholar
  3. 3.
    Bingle L, Brown NJ, Lewis CE (2002) The role of tumour-associated macrophages in tumour progression: implications for new anticancer therapies. J Pathol 196:254–265PubMedCrossRefGoogle Scholar
  4. 4.
    Chilov D, Kukk E, Taira S, Jeltsch M, Kaukonen J, Palotie A, Joukov V, Alitalo K (1997) Genomic organization of human and mouse genes for vascular endothelial growth factor C. J Biol Chem 272:25176–25183PubMedCrossRefGoogle Scholar
  5. 5.
    Clarijs R, Ruiter DJ, de Waal RM (2001) Lymphangiogenesis in malignant tumours: does it occur. J Pathol 193:143–146PubMedCrossRefGoogle Scholar
  6. 6.
    Cueni LN, Detmar M (2006) New insights into the molecular control of the lymphatic vascular system and its role in disease. J Invest Dermatol 126:2167–2177PubMedCrossRefGoogle Scholar
  7. 7.
    Dadras SS, Lange-Asschenfeldt B, Velasco P, Nguyen L, Vora A, Muzikansky A, Jahnke K, Hauschild A, Hirakawa S, Mihm MC, Detmar M (2005) Tumor lymphangiogenesis predicts melanoma metastasis to sentinel lymph nodes. Mod Pathol 18:1232–1242PubMedCrossRefGoogle Scholar
  8. 8.
    Dadras SS, Paul T, Bertoncini J, Brown LF, Muzikansky A, Jackson DG, Ellwanger U, Garbe C, Mihm MC, Detmar M (2003) Tumor lymphangiogenesis: a novel prognostic indicator for cutaneous melanoma metastasis and survival. Am J Pathol 162:1951–1960PubMedGoogle Scholar
  9. 9.
    Enholm B, Karpanen T, Jeltsch M, Kubo H, Stenback F, Prevo R, Jackson DG, Yla-Herttuala S, Alitalo K (2001) Adenoviral expression of vascular endothelial growth factor-C induces lymphangiogenesis in the skin. Circ Res 88:623–629PubMedGoogle Scholar
  10. 10.
    Esposito I, Menicagli M, Funel N, Bergmann F, Boggi U, Mosca F, Bevilacqua G, Campani D (2004) Inflammatory cells contribute to the generation of an angiogenic phenotype in pancreatic ductal adenocarcinoma. J Clin Pathol 57:630–636PubMedCrossRefGoogle Scholar
  11. 11.
    Feng HC, Song YF, Wen YM (2004) [Correlation between expression of vascular endothelial growth factor-C in tumor-associated macrophages and lymphatic metastasis in oral cancer]. Ai Zheng 23:278–281PubMedGoogle Scholar
  12. 12.
    Gershenwald JE, Thompson W, Mansfield PF, Lee JE, Colome MI, Tseng CH, Lee JJ, Balch CM, Reintgen DS, Ross MI (1999) Multi-institutional melanoma lymphatic mapping experience: the prognostic value of sentinel lymph node status in 612 stage I or II melanoma patients. J Clin Oncol 17:976–983PubMedGoogle Scholar
  13. 13.
    Goldman J, Le TX, Skobe M, Swartz MA (2005) Overexpression of VEGF-C causes transient lymphatic hyperplasia but not increased lymphangiogenesis in regenerating skin. Circ Res 96:1193–1199PubMedCrossRefGoogle Scholar
  14. 14.
    He Y, Karpanen T, Alitalo K (2004) Role of lymphangiogenic factors in tumor metastasis. Biochim Biophys Acta 1654:3–12PubMedGoogle Scholar
  15. 15.
    He Y, Kozaki K, Karpanen T, Koshikawa K, Yla-Herttuala S, Takahashi T, Alitalo K (2002) Suppression of tumor lymphangiogenesis and lymph node metastasis by blocking vascular endothelial growth factor receptor 3 signaling. J Natl Cancer Inst 94:819–825PubMedGoogle Scholar
  16. 16.
    Hirakawa S, Brown LF, Kodama S, Paavonen K, Alitalo K, Detmar M (2007) VEGF-C-induced lymphangiogenesis in sentinel lymph nodes promotes tumor metastasis to distant sites. Blood 109:1010–1017PubMedCrossRefGoogle Scholar
  17. 17.
    Jackowski S, Janusch M, Fiedler E, Marsch WC, Ulbrich EJ, Gaisbauer G, Dunst J, Kerjaschki D, Helmbold P (2007) Radiogenic lymphangiogenesis in the skin. Am J Pathol 171:338–348PubMedCrossRefGoogle Scholar
  18. 18.
    Jeltsch M, Kaipainen A, Joukov V, Meng X, Lakso M, Rauvala H, Swartz M, Fukumura D, Jain RK, Alitalo K (1997) Hyperplasia of lymphatic vessels in VEGF-C transgenic mice. Science 276:1423–1425PubMedCrossRefGoogle Scholar
  19. 19.
    Joukov V, Pajusola K, Kaipainen A, Chilov D, Lahtinen I, Kukk E, Saksela O, Kalkkinen N, Alitalo K (1996) A novel vascular endothelial growth factor, VEGF-C, is a ligand for the Flt4 (VEGFR-3) and KDR (VEGFR-2) receptor tyrosine kinases. EMBO J 15:1751PubMedGoogle Scholar
  20. 20.
    Joukov V, Sorsa T, Kumar V, Jeltsch M, Claesson-Welsh L, Cao Y, Saksela O, Kalkkinen N, Alitalo K (1997) Proteolytic processing regulates receptor specificity and activity of VEGF-C. EMBO J 16:3898–3911PubMedCrossRefGoogle Scholar
  21. 21.
    Karkkainen MJ, Haiko P, Sainio K, Partanen J, Taipale J, Petrova TV, Jeltsch M, Jackson DG, Talikka M, Rauvala H, Betsholtz C, Alitalo K (2004) Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins. Nat Immunol 5:74–80PubMedCrossRefGoogle Scholar
  22. 22.
    Karpanen T, Egeblad M, Karkkainen MJ, Kubo H, Yla-Herttuala S, Jaattela M, Alitalo K (2001) Vascular endothelial growth factor C promotes tumor lymphangiogenesis and intralymphatic tumor growth. Cancer Res 61:1786–1790PubMedGoogle Scholar
  23. 23.
    Kukk E, Lymboussaki A, Taira S, Kaipainen A, Jeltsch M, Joukov V, Alitalo K (1996) VEGF-C receptor binding and pattern of expression with VEGFR-3 suggests a role in lymphatic vascular development. Development 122:3829–3837PubMedGoogle Scholar
  24. 24.
    Landi G, Polverelli M, Moscatelli G, Morelli R, Landi C, Fiscelli O, Erbazzi A (2000) Sentinel lymph node biopsy in patients with primary cutaneous melanoma: study of 455 cases. J Eur Acad Dermatol Venereol 14:35–45PubMedCrossRefGoogle Scholar
  25. 25.
    Lee J, Gray A, Yuan J, Luoh SM, Avraham H, Wood WI (1996) Vascular endothelial growth factor-related protein: a ligand and specific activator of the tyrosine kinase receptor Flt4. Proc Natl Acad Sci U S A 93:1988–1992PubMedCrossRefGoogle Scholar
  26. 26.
    Lin J, Lalani AS, Harding TC, Gonzalez M, Wu WW, Luan B, Tu GH, Koprivnikar K, VanRoey MJ, He Y, Alitalo K, Jooss K (2005) Inhibition of lymphogenous metastasis using adeno-associated virus-mediated gene transfer of a soluble VEGFR-3 decoy receptor. Cancer Res 65:6901–6909PubMedCrossRefGoogle Scholar
  27. 27.
    Makinen T, Veikkola T, Mustjoki S, Karpanen T, Catimel B, Nice EC, Wise L, Mercer A, Kowalski H, Kerjaschki D, Stacker SA, Achen MG, Alitalo K (2001) Isolated lymphatic endothelial cells transduce growth, survival and migratory signals via the VEGF-C/D receptor VEGFR-3. EMBO J 20:4762–4773PubMedCrossRefGoogle Scholar
  28. 28.
    Mandriota SJ, Jussila L, Jeltsch M, Compagni A, Baetens D, Prevo R, Banerji S, Huarte J, Montesano R, Jackson DG, Orci L, Alitalo K, Christofori G, Pepper MS (2001) Vascular endothelial growth factor-C-mediated lymphangiogenesis promotes tumour metastasis. EMBO J 20:672–682PubMedCrossRefGoogle Scholar
  29. 29.
    Massi D, Puig S, Franchi A, Malvehy J, Vidal-Sicart S, Gonzalez-Cao M, Baroni G, Ketabchi S, Palou J, Santucci M (2006) Tumour lymphangiogenesis is a possible predictor of sentinel lymph node status in cutaneous melanoma: a case–control study. J Clin Pathol 59:166–173PubMedCrossRefGoogle Scholar
  30. 30.
    Nicosia RF (1998) What is the role of vascular endothelial growth factor-related molecules in tumor angiogenesis. Am J Pathol 153:11–16PubMedGoogle Scholar
  31. 31.
    Oh SJ, Jeltsch MM, Birkenhager R, McCarthy JE, Weich HA, Christ B, Alitalo K, Wilting J (1997) VEGF and VEGF-C: specific induction of angiogenesis and lymphangiogenesis in the differentiated avian chorioallantoic membrane. Dev Biol 188:96–109PubMedCrossRefGoogle Scholar
  32. 32.
    Olofsson B, Jeltsch M, Eriksson U, Alitalo K (1999) Current biology of VEGF-B and VEGF-C. Curr Opin Biotechnol 10:528–535PubMedCrossRefGoogle Scholar
  33. 33.
    Paavonen K, Horelli-Kuitunen N, Chilov D, Kukk E, Pennanen S, Kallioniemi OP, Pajusola K, Olofsson B, Eriksson U, Joukov V, Palotie A, Alitalo K (1996) Novel human vascular endothelial growth factor genes VEGF-B and VEGF-C localize to chromosomes 11q13 and 4q34, respectively. Circulation 93:1079–1082PubMedGoogle Scholar
  34. 34.
    Papoutsi M, Siemeister G, Weindel K, Tomarev SI, Kurz H, Schachtele C, Martiny-Baron G, Christ B, Marme D, Wilting J (2000) Active interaction of human A375 melanoma cells with the lymphatics in vivo. Histochem Cell Biol 114:373–385PubMedGoogle Scholar
  35. 35.
    Pepper MS (2001) Lymphangiogenesis and tumor metastasis: myth or reality? Clin Cancer Res 7:462–468PubMedGoogle Scholar
  36. 36.
    Saaristo A, Veikkola T, Enholm B, Hytonen M, Arola J, Pajusola K, Turunen P, Jeltsch M, Karkkainen MJ, Kerjaschki D, Bueler H, Yla-Herttuala S, Alitalo K (2002) Adenoviral VEGF-C overexpression induces blood vessel enlargement, tortuosity, and leakiness but no sprouting angiogenesis in the skin or mucous membranes. FASEB J 16:1041–1049PubMedCrossRefGoogle Scholar
  37. 37.
    Salven P, Lymboussaki A, Heikkila P, Jaaskela-Saari H, Enholm B, Aase K, von Euler G, Eriksson U, Alitalo K, Joensuu H (1998) Vascular endothelial growth factors VEGF-B and VEGF-C are expressed in human tumors. Am J Pathol 153:103–108PubMedGoogle Scholar
  38. 38.
    Schietroma C, Cianfarani F, Lacal PM, Odorisio T, Orecchia A, Kanitakis J, D’Atri S, Failla CM, Zambruno G (2003) Vascular endothelial growth factor-C expression correlates with lymph node localization of human melanoma metastases. Cancer 98:789–797PubMedCrossRefGoogle Scholar
  39. 39.
    Schoppmann SF, Birner P, Stockl J, Kalt R, Ullrich R, Caucig C, Kriehuber E, Nagy K, Alitalo K, Kerjaschki D (2002) Tumor-associated macrophages express lymphatic endothelial growth factors and are related to peritumoral lymphangiogenesis. Am J Pathol 161:947–956PubMedGoogle Scholar
  40. 40.
    Schoppmann SF, Fenzl A, Nagy K, Unger S, Bayer G, Geleff S, Gnant M, Horvat R, Jakesz R, Birner P (2006) VEGF-C expressing tumor-associated macrophages in lymph node positive breast cancer: impact on lymphangiogenesis and survival. Surgery 139:839–846PubMedCrossRefGoogle Scholar
  41. 41.
    Shields JD, Borsetti M, Rigby H, Harper SJ, Mortimer PS, Levick JR, Orlando A, Bates DO (2004) Lymphatic density and metastatic spread in human malignant melanoma. Br J Cancer 90:693–700PubMedCrossRefGoogle Scholar
  42. 42.
    Shimizu K, Kubo H, Yamaguchi K, Kawashima K, Ueda Y, Matsuo K, Awane M, Shimahara Y, Takabayashi A, Yamaoka Y, Satoh S (2004) Suppression of VEGFR-3 signaling inhibits lymph node metastasis in gastric cancer. Cancer Sci 95:328–333PubMedCrossRefGoogle Scholar
  43. 43.
    Skobe M, Hamberg LM, Hawighorst T, Schirner M, Wolf GL, Alitalo K, Detmar M (2001) Concurrent induction of lymphangiogenesis, angiogenesis, and macrophage recruitment by vascular endothelial growth factor-C in melanoma. Am J Pathol 159:893–903PubMedGoogle Scholar
  44. 44.
    Skobe M, Hawighorst T, Jackson DG, Prevo R, Janes L, Velasco P, Riccardi L, Alitalo K, Claffey K, Detmar M (2001) Induction of tumor lymphangiogenesis by VEGF-C promotes breast cancer metastasis. Nat Med 7:192–198PubMedCrossRefGoogle Scholar
  45. 45.
    Stacker SA, Williams RA, Achen MG (2004) Lymphangiogenic growth factors as markers of tumor metastasis. APMIS 112:539–549PubMedCrossRefGoogle Scholar
  46. 46.
    Straume O, Jackson DG, Akslen LA (2003) Independent prognostic impact of lymphatic vessel density and presence of low-grade lymphangiogenesis in cutaneous melanoma. Clin Cancer Res 9:250–256PubMedGoogle Scholar
  47. 47.
    Takizawa H, Kondo K, Fujino H, Kenzaki K, Miyoshi T, Sakiyama S, Tangoku A (2006) The balance of VEGF-C and VEGFR-3 mRNA is a predictor of lymph node metastasis in non-small cell lung cancer. Br J Cancer 95:75–79PubMedCrossRefGoogle Scholar
  48. 48.
    Thiele W, Sleeman JP (2006) Tumor-induced lymphangiogenesis: a target for cancer therapy? J Biotechnol 124:224–241PubMedCrossRefGoogle Scholar
  49. 49.
    Tobler NE, Detmar M (2006) Tumor and lymph node lymphangiogenesis—impact on cancer metastasis. J Leukoc Biol 80:691–696PubMedCrossRefGoogle Scholar
  50. 50.
    Valencak J, Heere-Ress E, Kopp T, Schoppmann SF, Kittler H, Pehamberger H (2004) Selective immunohistochemical staining shows significant prognostic influence of lymphatic and blood vessels in patients with malignant melanoma. Eur J Cancer 40:358–364PubMedCrossRefGoogle Scholar
  51. 51.
    Veikkola T, Jussila L, Makinen T, Karpanen T, Jeltsch M, Petrova TV, Kubo H, Thurston G, McDonald DM, Achen MG, Stacker SA, Alitalo K (2001) Signalling via vascular endothelial growth factor receptor-3 is sufficient for lymphangiogenesis in transgenic mice. EMBO J 20:1223–1231PubMedCrossRefGoogle Scholar
  52. 52.
    Wagner JD, Corbett L, Park HM, Davidson D, Coleman JJ, Havlik RJ, Hayes JT 2nd (2000) Sentinel lymph node biopsy for melanoma: experience with 234 consecutive procedures. Plast Reconstr Surg 105:1956–1966PubMedCrossRefGoogle Scholar
  53. 53.
    Was H (2005) [Characterization of markers and growth factors for lymphatic endothelium]. Postepy Biochem 51:209–214PubMedGoogle Scholar
  54. 54.
    Wissmann C, Detmar M (2006) Pathways targeting tumor lymphangiogenesis. Clin Cancer Res 12:6865–6868PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Barbara Boone
    • 1
  • Willeke Blokx
    • 2
  • Dirk De Bacquer
    • 3
  • Jo Lambert
    • 1
  • Dirk Ruiter
    • 2
  • Lieve Brochez
    • 1
  1. 1.Department of DermatologyUniversity Hospital GhentGhentBelgium
  2. 2.Department of PathologyUMC Sint-RadboudNijmegenThe Netherlands
  3. 3.Department of Public HealthUniversity Hospital GhentGhentBelgium

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