The lymphatic system plays an important role in both tissue homeostasis and inflammation. During the surgical procedure there is complete disruption of lymphatic drainage of the allograft kidney. The time course and nature of lymphatic reconnection following transplantation is poorly understood. In addition to the extra-renal lymphangiogenesis required for lymphatic reconnection, some patients may develop de novo lymphatic vessels within the renal parenchyma during acute rejection or chronic allograft damage. This work sought to examine the time course and mechanism of lymphangiogenesis and the role of macrophages in this process. Injection of carbon black and Evan’s blue into the rat kidney resulted in rapid transit to the draining hilar renal lymph node. Surgical disruption of the lymphatic drainage of the kidney prevented trafficking of carbon black to the renal lymph node at 24 hours. At day 6 there was macroscopic and microscopic evidence of carbon black localisation in the renal lymph node suggesting functional reconnection. Careful histological analysis of hilar renal tissue indicated that the large lymphatic trunks were replaced by a network of small proliferating lymphatic vessels. Assessment of intra-renal lymphangiogenesis was undertaken in 2 distinct experimental models of renal transplantation. In a murine model of acute allograft rejection there was no evidence of increased lymphatic vessel number at day 7. In a collaboration with Sheffield University, tissue from a rat model of interstitial fibrosis and tubular atrophy was examined. The rat tissue exhibited a prominent macrophage and T-cell infiltration at 12 months but there was no difference in the number of perivascular lymphatic vessels. In contrast, there were numerous lymphatic vessels evident in the interstitium that were absent in control isograft tissue. Interestingly, the number of lymphatic vessels correlated with the extent of fibrosis. Analysis of vascular endothelial cell growth factor-C (VEGF-C) mRNA expression did not show any increase in allografts. The model of unilateral ureteric obstruction (UUO) was employed as a model of rapidly progressive inflammatory fibrosis. UUO was associated with rapid and prominent interstitial lymphangiogenesis. This was associated with a marked increase in macrophage and T-lymphocyte infiltration and increased whole kidney mRNA expression of VEGF-C. The role of macrophages in lymphangiogenesis was explored by administration of macrophage depleting liposomal clodronate. No effect upon lymphangiogenesis was found but liposomal clodronate failed to deplete ED-1 positive macrophages in the kidney. A macrophage isolation strategy was thus employed using the myeloid CD11b marker cells and flow cytometric cell sorting and immunomagnetic bead sorting. Although gene expression studies demonstrated increased ED1 mRNA expression by CD11b enriched cells, no difference in VEGF-C mRNA expression between CD11b cells obtained from obstructed kidneys versus cells from sham controls was evident. Lastly, despite extensive efforts, immunostaining for VEGF-C was unsuccessful. In summary, lymphangiogenesis can reconstitute the lymphatic drainage of the kidney and is prominent in both chronic allograft injury and the acutely obstructed kidney in the rat. Although VEGF-C is the likely driver of lymphangiogenesis direct evidence of macrophage VEGF-C production was not found.

The lymphatic vasculature is a network of unidirectional capillaries and ducts, which serve to return extracellular fluid and macromolecules to the systemic blood circulation. In addition, the lymphatic vessels have important roles in immune surveillance and fat absorption. Dysfunction of lymphatic vessels has profound physiological consequences. Insufficient lymph uptake results in lymphedema, a chronic disabling condition that currently has no cure. Lymphedemcanoccureither due to developmental defects (primary lymphedema) or due to injuries of existing lymphatic vessels (secondary lymphedema). The significance of lymphangiogenesis in tumour metastasis is demonstrated by the finding that increased lymphangiogenesis is associated with a higher rate of metastasis and poorer prognosis in cancer patients. Prox1, a homeoboxgene, regulates the development of the lymphatic vasculature by upregulating the expression of lymphatic endothelial markers and simultaneously repressing the expression of blood endothelial markers. To explore the mechanisms by which Prox1 establishes lymphatic cell fate, we compared Prox1 mediated activation oftwo key lymphatic cell surface receptors: theVascular Endothelial Growth Factor Receptor-3 (VEGFR-3) and theFibroblast Growth Factor Receptor-3 (FGFR-3) genes. Using a combination of luciferase gene reporter assays and immunoblotting, we compared the ability of different Prox1 constructs to activate either VEGFR-3 or FGFR-3 at both the mRNA and protein levels, respectively. Furthermore, we tested whether recombinant PROX1 protein was able to bind to the proximal promoter regions of VEGFR-3 and FGFR-3usingelectrophoretic mobility shift assays (EMSA). DNA binding deficient Prox1 versions did not activate the FGFR-3 promoter. In contrast, these versions of Prox1still efficiently activated transcription of the VEGFR-3 promoter. In agreement with our luciferase reporter gene assays, immunoblotting of HUVECs demonstrated that only infection with wt Prox1adenovirus increased expression of the FGFR-3 protein. Infection of HUVECs with adenoviral vectors encoding eitherwt Prox1orHDPD∆ Prox1was sufficient to induce a significant increase in VEGFR-3 protein levels. Surprisingly, our EMSA results with recombinant PROX1 demonstrated that PROX1 can bind to the promoter region of both VEGFR-3 and FGFR-3 genes via its DNA binding domain. We showed that PROX1 potentially binds to the promoter region of the VEGFR-3 gene via a consensus Prospero binding site (CGCCTCGGC). Our data demonstrates that, in endothelial cells, PROX1 utilizes distinct mechanisms to activate these two key endothelial growth factor receptors.

The lymphatic vasculature is a network of unidirectional capillaries and ducts, which serve to return extracellular fluid and macromolecules to the systemic blood circulation. In addition, the lymphatic vessels have important roles in immune surveillance and fat absorption. Dysfunction of lymphatic vessels has profound physiological consequences. Insufficient lymph uptake results in lymphedema, a chronic disabling condition that currently has no cure. Lymphedemcanoccureither due to developmental defects (primary lymphedema) or due to injuries of existing lymphatic vessels (secondary lymphedema). The significance of lymphangiogenesis in tumour metastasis is demonstrated by the finding that increased lymphangiogenesis is associated with a higher rate of metastasis and poorer prognosis in cancer patients. Prox1, a homeoboxgene, regulates the development of the lymphatic vasculature by upregulating the expression of lymphatic endothelial markers and simultaneously repressing the expression of blood endothelial markers. To explore the mechanisms by which Prox1 establishes lymphatic cell fate, we compared Prox1 mediated activation oftwo key lymphatic cell surface receptors: theVascular Endothelial Growth Factor Receptor-3 (VEGFR-3) and theFibroblast Growth Factor Receptor-3 (FGFR-3) genes. Using a combination of luciferase gene reporter assays and immunoblotting, we compared the ability of different Prox1 constructs to activate either VEGFR-3 or FGFR-3 at both the mRNA and protein levels, respectively. Furthermore, we tested whether recombinant PROX1 protein was able to bind to the proximal promoter regions of VEGFR-3 and FGFR-3usingelectrophoretic mobility shift assays (EMSA). DNA binding deficient Prox1 versions did not activate the FGFR-3 promoter. In contrast, these versions of Prox1still efficiently activated transcription of the VEGFR-3 promoter. In agreement with our luciferase reporter gene assays, immunoblotting of HUVECs demonstrated that only infection with wt Prox1adenovirus increased expression of the FGFR-3 protein. Infection of HUVECs with adenoviral vectors encoding eitherwt Prox1orHDPD∆ Prox1was sufficient to induce a significant increase in VEGFR-3 protein levels. Surprisingly, our EMSA results with recombinant PROX1 demonstrated that PROX1 can bind to the promoter region of both VEGFR-3 and FGFR-3 genes via its DNA binding domain. We showed that PROX1 potentially binds to the promoter region of the VEGFR-3 gene via a consensus Prospero binding site (CGCCTCGGC). Our data demonstrates that, in endothelial cells, PROX1 utilizes distinct mechanisms to activate these two key endothelial growth factor receptors.

Le psoriasis est une pathologie cutanée chronique fréquente qui semble déterminée par des facteurs génétiques multiples et par des facteurs environnementaux. L'expansion vasculaire est un processus précoce lors du développement des plaques de psoriasis. La lymphangiogenèse a jusqu'alors été peu étudiée dans le développement de ces lésions bien que l'expansion du réseau lymphatique y soit histologiquement décrite. Des anomalies dans les systèmes régulant ces processus pourraient faire partie de la pathogénie du psoriasis. Nous avons étudié les réseaux vasculaires et lymphatiques ainsi que lexpression de facteurs régulant leur expansion dans le psoriasis vulgaire en plaques. Nos résultats montrent une surexpression de l'ARNm du VEGF-A 121 et du VEGFR3 dans la peau non lésionnelle des patients atteints de psoriasis sans altération histologique décelable par rapport à la peau de volontaires sains. En outre, l'étude de lésions débutantes de type pinpoint en comparaison des plaques établies de psoriasis montre que l'expansion lymphatique suit l'expansion vasculaire sanguine dans le psoriasis. Les facteurs VEGFR3 et prox-1, marqueurs des vaisseaux lymphatiques, sont en effet exprimés de manière intermédiaire dans les pinpoint par rapport aux plaques et à la peau saine contrairement aux marqueurs d'angiogenèse, déjà exprimés de manière identique entre pinpoint et plaques. Le VEGF-A 189 est accru dans les lésions débutantes, ce qui suggère un rôle pour cette isoforme dans l'élongation vasculaire qui accompagne le psoriasis. En outre, la régression des lésions au cours d'un traitement montre que le réseau lymphatique régresse après le réseau sanguin. Ce travail démontre des différences dans le transcriptome vasculaire (sanguin et lymphatique) de la peau saine des patients par rapport à celle des volontaires sains et une chronologie séquentielle pour le développement sanguin et lymphatique dans le psoriasis. Il met aussi en évidence les variations d'expression de facteurs contrôlant l'angiogenèse et la lymphangiogenèse dans diverses lésions de psoriasis par rapport à la peau saine.

Inflammatory cytokines and chemokines are known to promote tumor cell survival, invasion, the formation of blood and lymphatic vessels, and hence, metastasis. We previously showed that a pro-inflammatory pathway regulated by Toll-like Receptor-4 (TLR4) can be activated in human breast cancer (BC) cell lines by a clinical chemotherapeutic drug, paclitaxel (PXL). Prior data showed that PXL treatment of TLR4+ tumors in vivo increased inflammation and tumor spread. Here, we used two BC models based on MDA-MB-231 and HCC1806 cell lines. Transcript expression of 123 cytokines in vitro and in vivo was determined by qRT-PCR. We found that 18 and 26 cytokines were upregulated by nanoparticle albumin-bound paclitaxel (nab-PXL) in 231TLR4+ cells and in 1806TLR4+ cells, respectively. Upregulation of cytokines was observed in cultured cells and in tumor models in vivo. Furthermore, fourteen cytokines (11.3% of total) were induced by nab-PXL in both tumor models suggesting that these targets are upregulated by PXL regardless of genetic makeup of tumor cells. We also confirmed the expression of these cytokines on protein level by ELISA. We found that expression of CCL20 and CXCL1 proteins is consistent with transcript expression detected by qRT-PCR. Taken together, CCL20 and CXCL1 can be the potential targets for further study, and they may have the capacity to recruit myeloid-derived lymphatic endothelial cell progenitors to the tumor site to promote lymphangiogenesis in breast cancer.

Το αδενοκαρκίνωμα του προστάτη αποτελεί μείζων και κλιμακούμενο πρόβλημα παγκοσμίως. Σε πολλές ανεπτυγμένες χώρες ο καρκίνος του προστάτη αποτελεί την πιο κοινά διαγνωσμένη κακοήθεια στους άνδρες. Η μετάσταση των καρκινικών κυττάρων αποτελεί την κύρια αιτία θνησιμότητας και συχνά συμβαίνει μέσω του λεμφαγγειακού συστήματος που αποτελεί την κυριότερη οδό για την διαφυγή των καρκινικών κυττάρων. Η εξάπλωση των καρκινικών κυττάρων στους λεμφαδένες είναι ένα από τα πρώτα βήματα της μεταστατικής διαδικασίας και αποτελεί καθοριστικό παράγοντα για την πρόγνωση. Η λεμφαγγειογένεση (σχηματισμός νέων λεμφαγγείων) θεωρείται καθοριστική για την μετάσταση των καρκινικών κυττάρων στους λεμφαδένες. Οι λεμφαγγειογενετικοί παράγοντες φαίνεται να προάγουν, τη δημιουργία νέων λεμφαγγείων καθώς και τη μετάσταση του πρωτοπαθούς όγκου στους λεμφαδένες. Ένα από τους παράγοντες αυτούς είναι ο VEGF-C. Σκοπός. Η μελέτη αυτή σχεδιάστηκε για την σύγκριση της λεμφαγγειοβρίθειας μεταξύ προστατικών καρκινωμάτων με και χωρίς λεμφαδενικές μεταστάσεις. Σκοπό της εργασίας αποτελούσε επίσης η διευκρίνιση του ρόλου που παίζει ο λεμφαγγειογενετικός παράγοντας VEGF-C στις λεμφαδενικές μεταστάσεις. Σχεδιασμός της μελέτης. Για τον λόγο αυτό εγκλεισμένοι σε παραφίνη ιστοί 87 ασθενών οι οποίοι υποβλήθηκαν σε ριζική προστατεκτομή επεξεργάσθηκαν με τη διαδικασία της ανοσοϊστοχημείας με μονοκλωνικά αντισώματα για της πρωτεΐνες CD-31, Lyve-1, podoplanin και VEGF-C. Αποτελέσματα. Ετερογενής χρώση για τον VEGF-C παρατηρήθηκε στο σύνολο των καρκινικών δειγμάτων. Η ανοσοχρώση παρουσίαζε μεγαλύτερη ένταση στα περιστατικά με λεμφαδενικές μεταστάσεις σε σχέση με αυτά που έφεραν αρνητικούς λεμφαδένες. Μεγαλύτερη λεμφαγγειοβρίθεια εντοπίσθηκε γύρω από τον όγκο σε σχέση με τις εντός του όγκου περιοχές. Η λεμφαγγειοβρίθεια δεν σχετίσθηκε με την ύπαρξη μεταστάσεων ούτε με το grade του όγκου. Συμπεράσματα. Από κοινού τα αποτελέσματα αυτά προτείνουν ότι η λεμφαγγειοβρίθεια δεν μπορεί να αποτελέσει προγνωστική μεταβλητή των λεμφαδενικών μεταστάσεων στο αδενοκαρκίνωμα του προστάτη. Ο VEGF-C παρουσιάζει έντονη έκφραση στις καρκινικές περιοχές και η σχέση που εντοπίσθηκε με τις λεμφαδενικές μεταστάσεις προτείνει ένα πιθανό ρόλο στην ανάπτυξη λεμφαδενικών μεταστάσεων. Πχ μέσω της αυξημένης διαπερατότητας των τοιχωμάτων των λεμφαγγείων. / Carcinoma of the prostate now constitutes a major and escalating international health problem. In many developed countries prostate cancer is the most commonly diagnosed malignancy in men. The metastatic spread of tumor cells is the most lethal aspect of cancer and often occurs via the lymphatic vasculature which serves as a major route for tumour metastasis. The dissemination of malignant cells to the regional lymph nodes is an early step in the progression of prostate tumours and is an important determinant of prognosis. Lymphangiogenesis (formation of new lymphatic vessels) is thought to be crucial for cancer cells to metastasise to the regional lymph nodes. Lymphangiogenic growth factors have been identified that promote formation of tumor lymphatics and metastatic spread of tumor cells to lymph nodes. These include the secreted glycoproteins vascular endothelial growth factor- C (VEGF-C) Purpose: This study was designed to compare prostate carcinoma, with and with no lymph node metastasis, for lymphatic vessel density (LVD) and the expression of the lymph-endothelial specific growth factor, vascular endothelial growth factor C (VEGF-C), to determine their role in lymphogenic metastasis. Experimental Design: For this purpose paraffin-embedded tissues obtained from 87 patients who underwent radical prostatectomy were immunostained with D2-40, CD-31, Lyve-1 and VEGF-C monoclonal antibodies. The associations between the results were analyzed with statistic analysis protocols. Results: Widespread, heterogeneous staining for VEGF-C was observed in all cancer specimens. Intensity of VEGF-C staining was higher in carcinomas with lymph node metastasis than in those with negative lymph nodes. The highest LVD was found in periphery as opposed to tumorous areas. There was no correlation with clinical staging. Conclusions: Together, these results suggest that LVD is not a prognostic variable for the process of lymphogenic metastasis in prostate cancer. VEGF-C is up-regulated in prostate cancer and its correlation with lymph node status suggests a role for the development of lymph node metastasis. e.g. via an increased permeability of lymphatic vessels.

The generation of new lymphatic vessels through lymphangiogenesis has been implicated in many disease states. This process has some overlap with the better studied angiogenesis pathway, but is under distinct molecular control. Specifically, it has been shown that VEGFR-3 and neuropilin-2 are important mediators of lymphangiogenesis. A greater understanding of this process could lead to new therapies for cancer and lymphedemas. We investigated lymphatic vessel growth in a mouse model with a focus on the effects of neuropilin-2 knockout. First, we induced an immunogenic response via delayed-type hypersensitivity to examine lymphangiogenesis in the physiologic state. Our neuropilin-2 knockout mouse model displayed a decreased ability to resolve inflammation on exposure to an allergen. Next, we subcutaneously injected a highly invasive melanoma to examine lymphangiogenesis in the pathologic state. We noted significantly reduced tumor growth in our neuropilin-2 knockout. In addition, the neuropilin-2 knockout mice displayed reduced vessel area in comparison to their wild-type littermates, suggesting that inhibition of neuropilin-2 may prove a potent antitumor therapeutic strategy. These results highlight neuropilin-2's important role as a mediator of physiological and pathological angiogenesis and lymphangiogenesis.

ALK1 (Actlivin receptor-Like Kinase 1) est un récepteur exprimé spécifiquement sur les cellules endothéliales. ALK1 possède deux ligands de haute affinité BMP9 (Bone Morphogenic Protein) et BMP10. Cette voie de signalisation joue des rôles cruciaux dans l'angiogenèse et la lymphangiogenèse qui sont des processus critiques dans le développement mais aussi le cancer. Ainsi, l'objectif de cette thèse était de caractériser les fonctions de BMP9 et BMP10 dans le développement vasculaire physiologique et tumoral à l'aide de souris invalidées pour ces deux facteurs.Nous avons montré que BMP9 et BMP10 sont essentiels à la fermeture du canal artériel. Ce vaisseau permet de dévier le sang hors des poumons foetaux non fonctionnels et sa fermeture au moment de la naissance est nécessaire à la survie postnatale. L'absence de BMP9 et BMP10 mène à une réouverture du canal artériel chez le souriceau du fait de problèmes de remodelage. Chez l'Homme, une région critique contenant Bmp10 peut être corrélée à des problèmes de fermeture du canal artériel.Des essais cliniques ciblant ALK1 dans le traitement du cancer sont en cours du fait de son implication dans l'angiogenèse. Néanmoins, les rôles spécifiques de BMP9, BMP10 et ALK1 dans le cancer sont encore mal connus. Nous avons montré que BMP10 n'est pas impliqué dans la tumorigenèse mammaire au contraire de BMP9 qui agit comme un facteur de quiescence et de maturation dans l'angiogenèse tumorale. Ainsi, il serait plus intéressant de cibler spécifiquement BMP9 plutôt qu'ALK1 afin d'éviter d'altérer les fonctions physiologiques de BMP10. / ALK1 (Actlivin receptor-Like Kinase 1) is a receptor specifically expressed at the surface of endothelial cells. BMP9 (Bone Morphogenic Protein) and BMP10 are the high affinity ligands of ALK1. This pathway has been proved to play important roles in angiogenesis and lymphangiogenesis which are critical processes in development as well as in cancer. Thus, the purpose of my PhD was to characterize BMP9 and BMP10 functions in physiological and tumoral vascular development using mice invalidated for those proteins.We show that BMP9 and BMP10 are essentials to ductus arteriosus closure, a shunt allowing blood to avoid inoperative foetal lungs. Its closure is mandatory to survival after birth. Loss of BMP9 and BMP10 leads to ductus arteriosus reopening in mice pups due to remodeling issues. In human, a critical region including Bmp10 was correlated to patent ductus arteriosus.Clinical trials targeting ALK1 in cancer treatment are in progress due to its involvment in angiogenesis. However, little is known about the specific roles of BMP9, BMP10 and ALK1 in carcinogenesis. We show that BMP10 is not involved in mammary cancer development while BMP9 acts as a quiescent and maturation factor in tumoral angiogenesis. Thus, it may be more interesting to target only BMP9 instead of ALK1 to avoid interferences with BMP10 physiological functions.

No description available.

A expressão do fator de crescimento endotelial vascular tipo C (VEGF-C) e de podoplanina pelas células malignas tem sido associada com a maior ocorrência de metástases regionais e/ou pior prognóstico para os pacientes com câncer de boca. O objetivo deste estudo foi avaliar o significado clinico da expressão de VEGF-C e podoplanina em 42 carcinomas espinocelulares (CEC) bem diferenciado de boca, com e sem comprometimento linfonodal, incluindo oito carcinomas verrucosos, tratados no Departamento de Cirurgia de Cabeça e Pescoço, do Hospital do Câncer A. C. Camargo, São Paulo, no período de 1980 a 2000. Os pacientes foram analisados quanto ao gênero, idade, cor, tabagismo, etilismo, localização do tumor primário, classificação pelo sistema TNM, tratamento, ocorrência de recidivas local e regional, metástase à distância, de segundo tumor primário, além da presença de infiltrações perineural, muscular, glandular e óssea e de embolizações vasculares. Analisou-se também o índice histopatológico de malignidade tumoral e as expressões imuno-histoquímica de VEGFC e de podoplanina pelas células malignas no front de invasão tumoral. A associação das expressões de VEGF-C e podoplanina com as variáveis estudadas foi calculada pelo teste quiquadrado. As probabilidades de sobrevida, acumuladas nos períodos de 5 e 10 anos calculadas pelo método de Kaplan-Meier e comparadas pelo teste de long-rank. A maioria dos CEC de boca, incluindo os carcinomas verrucosos, exibiu uma forte expressão de VEGF-C pelas células malignas. Houve uma tendência, sem associação estatisticamente significativa, dos pacientes com CEC de boca e forte expressão de VEGF-C apresentarem maior freqüência de recidivas locais e/ou regionais. A forte expressão de podoplanina foi significativamente associada com o gênero masculino (p=0,037), com o estadiamento T1 e T2 (p=0,037), com o estadio clínico I e II (p=0,027) e com a presença de infiltração glandular (p=0,003). As recidivas locais e regionais foram detectadas mais freqüentemente nos CEC de boca com forte expressão de podoplanina, porém sem diferença estatisticamente significativa, quando comparados com aqueles com expressão fraca da proteína. As taxas de sobrevida global e específica por câncer para os pacientes com tumores com forte expressão de VEGF-C e a taxa de sobrevida global para a forte expressão de podoplanina foram percentualmente menores do que aquelas dos pacientes com tumores com fraca expressão destas proteínas. O comprometimento linfonodal cervical se mostrou fator de prognóstico significativo (p=0,001) para os pacientes com câncer de boca. Estes resultados sugerem que a forte expressão de VEGF-C e podoplanina pelas células malignas juntamente com o comprometimento linfonodal regional são fatores indicativos de uma evolução clínica desfavorável e de um pior prognóstico para os pacientes com CEC bem diferenciados de boca. / The expression of vascular endothelial growth factor C (VEGF-C) and podoplanin by malignant cells has been associated with a greater incidence of regional metastasis and/or poor prognosis for patients with oral cancer. The purpose of this study was to evaluate the clinical significance of VEGF-C and podoplanin expression in 42 well-differentiated oral squamous cell carcinomas (OSCC), with and without lymph node involvement, including eight verrucous carcinoma, treated at the Department of the Head and Neck Surgery and Otorhinolaryngology, A. C. Camargo Cancer Hospital, São Paulo, from 1980 to 2000. Patients were evaluated according the following parameters: gender, age, ethnic group, tobacco and/or alcohol consumption, tumor location, TNM stage, treatment and clinical follow-up (recurrence, occurrence of a second primary tumor and death) and the presence of perineural, muscular, glandular and bone infiltrations or vascular embolizations. In addition, we investigated the histopathological malignancy index and the immunohistochemistry expression of VEGF-C and podoplanin by malignant cells in the invasive front tumor. Chisquare test or Fishers exact test was used to analyze the association of VEGF-C and podoplanin with demographic, clinical and microscopic variables in OSCC patients. The 5 and 10-year survival rates were calculated by Kaplan-Meier method and the comparison of the survival curves were performed using log rank test. Most of OSCC, including verrucous carcinoma, showed a high expression of VEGF-C by malignant cells. The OSCC patients with high expression of VEGF-C showed a tendency, without statistical significance, of local and/or regional recurrence. The overexpression of podoplanin was significantly associated with the male gender (p=0,037), with T1 and T2 stage (p=0,037), with I and II clinical stage (p=0,027) and with the presence of glandular infiltration (p=0,003). The local and regional recurrences were detected more frequently in OSCC with high expression of podoplanin, without statistical significant difference, when compared with those with low expression of the protein. The overall survival rates and cancer specific survival rates for OSCC patients with high VEGF-C expression and the overall survival rate for OSCC patients with podoplanin overexpression were lower than those of OSCC patients with low expression of these proteins. The cervical lymph node involvement was significant prognostic factor (p=0,001) for patients with oral cancer. These results suggest that the overexpression of VEGF-C and podoplanin by malignant cells and regional lymph node involvement are indicative factors of an unfavorable clinical outcome and a poor prognosis for patients with well-differentiated OSCC.