Immunohistochemical Analysis of VEGF-C/VEGFR-3 System and Lymphatic Vessel Extent in Normal and Adenomatous Human Pituitary Tissues

Abstract

The angiogenic growth factor Vascular Endothelial Growth Factor-C (VEGF-C) and its receptor VEGFR-3 are also known to be implicated in the development of lymphatic vessels. We assessed the expression of VEGF-C and VEGFR-3, together with blood and lymphatic vessel extents and proliferation index (PI) values, by immunohistochemistry (IHC) in 6 normal human pituitary glands and 53 pituitary adenomas of different tumour grade, on consecutive tissue sections. VEGF-C was detected in around 10% of the endocrine cells in normal pituitary tissue, while this gland was devoid of lymphatic vascularization and showed very few vessels positive for VEGFR-3. Concerning tumour tissue, most of the adenomas showing VEGF-C immunoreactivity (21/47) were positive in 60% of the tumour cells and the ones positive for VEGFR-3 showed a number of immunostained vessels higher than those observed in the normal pituitary. Most of the tumours positive for VEGFR-3 did not show any LYVE-1 positive vessels (18/53), suggesting that at least in these cases, VEGFR-3 is expressed on blood vessels. Nevertheless, we observed a significant association between low expression of VEGFR-3 and low lymphatic vessel number, suggesting that VEGFR-3 might be involved in the starting of de novo lymphangiogenesis in this tumour type. Moreover, tumours bearing lymphatic vessels showed the tendency to shift towards a more aggressive behaviour (high tumour grade and high PI). In conclusion, the VEGF-C/VEGFR-3 system might be involved in controlling tumour angiogenesis in the pituitary adenomas lacking lymphatic vessels, but may also play a role in starting the process of tumour lymphangiogenesis.

Key words

VEGF-C - VEGFR-3 - pituitary adenomas - LYVE-1 - lymphatic vessels

References

• 1 Arinaga M, Noguchi T, Takeno S, Chujo M, Miura T, Uchida Y. Clinical significance of vascular endothelial growth factor C and vascular endothelial growth factor receptor 3 in patients with nonsmall cell lung carcinoma.  Cancer. 2003;  97 457-464
• 2 Banerji S, Ni J, Wang SX, Clasper S, Su J, Tammi R, Jones M, Jackson DG. LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.  J Cell Biol. 1999;  144 789-801
• 3 Bates AS, Farrell WE, Bicknell EJ, McNicol AM, Talbot AJ, Broome JC, Perrett CW, Thakker RV, Clayton RN. Allelic deletion in pituitary adenomas reflects aggressive biological activity and has potential value as a prognostic marker.  J Clin Endocrinol Metab. 1997;  82 818-824
• 4 Bjorndahl MA, Cao R, Burton JB, Brakenhielm E, Religa P, Galter D, Wu L, Cao Y. Vascular endothelial growth factor-a promotes peritumoral lymphangiogenesis and lymphatic metastasis.  Cancer Res. 2005;  65 9261-9268
• 5 Cao R, Bjorndahl MA, Religa P, Clasper S, Garvin S, Galter D, Meister B, Ikomi F, Tritsaris K, Dissing S, Ohhashi T, Jackson DG, Cao Y. PDGF-BB induces intratumoral lymphangiogenesis and promotes lymphatic metastasis.  Cancer Cell. 2004;  6 333-345
• 6 Dadras SS, Paul T, Bertoncini J, Brown LF, Muzikansky A, Jackson DG, Ellwanger U, Garbe C, Mihm MC, Detmar M. Tumor lymphangiogenesis: a novel prognostic indicator for cutaneous melanoma metastasis and survival.  Am J Pathol. 2003;  162 1951-1960
• 7 Gershenwald JE, Fidler IJ. Targeting lymphatic metastasis.  Science. 2002;  296 1811-1812
• 8 Jeltsch M, Kaipainen A, Joukov V, Meng X, Lakso M, Rauvala H, Swartz M, Fukumura D, Jain RK, Alitalo K. Hyperplasia of lymphatic vessels in VEGF-C transgenic mice.  Science. 1997;  276 1423-1425
• 9 Jüttner S, Wissmann C, Jons T, Vieth M, Hertel J, Gretschel S, Schlag PM, Kemmner W, Hocker M. Vascular endothelial growth factor-D and its receptor VEGFR-3: two novel independent prognostic markers in gastric adenocarcinoma.  J Clin Oncol. 2006;  24 228-240
• 10 Kaipainen A, Korhonen J, Mustonen T, Hinsbergh VW van, Fang GH, Dumont D, Breitman M, Alitalo K. Expression of the fms-like tyrosine kinase 4 gene becomes restricted to lymphatic endothelium during development.  Proc Natl Acad Sci U S A. 1995;  92 3566-3570
• 11 Kaltsas GA, Nomikos P, Kontogeorgos G, Buchfelder M, Grossman AB. Diagnosis and Management of Pituitary Carcinomas.  J Clin Endocrinol Metab. 2005;  90 3089-3099
• 12 Karkkainen MJ, Makinen T, Alitalo K. Lymphatic endothelium: a new frontier of metastasis research.  Nat Cell Biol. 2002;  4 2-5
• 13 Kubo H, Fujiwara T, Jussila L, Hashi H, Ogawa M, Shimizu K, Awane M, Sakai Y, Takabayashi A, Alitalo K, Yamaoka Y, Nishikawa SI. Involvement of vascular endothelial growth factor receptor-3 in maintenance of integrity of endothelial cell lining during tumor angiogenesis.  Blood. 2000;  96 546-553
• 14 Lohrer P, Gloddek J, Hopfner U, Losa M, Uhl E, Pagotto U, Stalla GK, Renner U. Vascular endothelial growth factor production and regulation in rodent and human pituitary tumor cells in vitro.  Neuroendocrinology. 2001;  74 95-105
• 15 Luzi P, Miracco C, Lio R, Malandrini A, Piovani S, Giovanni Venezia S, Tosi P. Endocrine inactive pituitary carcinoma metastasizing to cervical lymph nodes: a case report.  Hum Pathol. 1987;  18 90-92
• 16 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. Isolated lymphatic endothelial cells transduce growth, survival and migratory signals via the VEGF-C/D receptor VEGFR-3.  EMBO J. 2001;  20 4762-4773
• 17 Morisada T, Oike Y, Yamada Y, Urano T, Akao M, Kubota Y, Maekawa H, Kimura Y, Ohmura M, Miyamoto T, Nozawa S, Koh GY, Alitalo K, Suda T. Angiopoietin-1 promotes LYVE-1-positive lymphatic vessel formation.  Blood. 2005;  105 4649-4656
• 18 Mountcastle RB, Roof BS, Mayfield RK, Mordes DB, Sagel J, Biggs PJ, Rawe SE. Pituitary adenocarcinoma in an acromegalic patient: response to bromocriptine and pituitary testing: a review of the literature on 36 cases of pituitary carcinoma.  Am J Med Sci. 1989;  298 109-118
• 19 Oh SJ, Jeltsch MM, Birkenhager R, McCarthy JE, Weich HA, Christ B, Alitalo K, Wilting J. VEGF and VEGF-C: specific induction of angiogenesis and lymphangiogenesis in the differentiated avian chorioallantoic membrane.  Dev Biol. 1997;  188 96-109
• 20 Onofri C, Theodoropoulou M, Losa M, Uhl E, Lange M, Arzt E, Stalla GK, Renner U. Localization of vascular endothelial growth factor (VEGF) receptors in normal and adenomatous pituitaries: detection of a non-endothelial function of VEGF in pituitary tumours.  J Endocrinol. 2006;  191 249-261
• 21 Padera TP, Kadambi A, Thomaso E Di, Carreira CM, Brown EB, Boucher Y. Lymphatic metastasis in the absence of functional intra tumour lymphatics.  Science. 2002;  296 1883-1886
• 22 Partanen TA, Arola J, Saaristo A, Jussila L, Ora A, Miettinen M, Stacker SA, Achen MG, Alitalo K. VEGF-C and VEGF-D expression in neuroendocrine cells and their receptor, VEGFR-3, in fenestrated blood vessels in human tissues.  FASEB J. 2000;  14 2087-2096
• 23 Stacker SA, Caesar C, Baldwin ME, Thornton GE, Williams RA, Prevo R, Jackson DG, Nishikawa S, Kubo H, Achen MG. VEGF-D promotes the metastatic spread of tumor cells via the lymphatics.  Nat Med. 2001;  7 186-191
• 24 Stacker SA, Baldwin ME, Achen MG. The role of tumor lymphangiogenesis in metastatic spread.  FASEB J. 2002;  16 922-934
• 25 Vidal S, Kovacs K, Horvath E, Scheithauer BW, Kuroki T, Lloyd RV. Microvessel density in pituitary adenomas and carcinomas.  Virchows Arch. 2001;  438 595-602
• 26 Walker JD, Grossman A, Anderson JV, Ur E, Trainer PJ, Benn J, Lowy C, Sonksen PH, Plowman PN, Lowe DG. Malignant prolactinoma with extracranial metastases: a report of three cases.  Clin Endocrinol (Oxf). 1993;  38 411-419
• 27 Yonemura Y, Fushida S, Bando E, Kinoshita K, Miwa K, Endo Y, Sugiyama K, Partanen T, Yamamoto H, Sasaki T. Lymphangiogenesis and the vascular endothelial growth factor receptor (VEGFR)-3 in gastric cancer.  Eur J Cancer. 2001;  37 918-923

Correspondence