• PD-1
• cytokines
• exhausted T cells
• squamous cell carcinoma
• tumor infiltrating lymphocytes
• tumor microenvironment

Nodal metastasis is a critical factor in predicting the prognosis of oral squamous cell carcinoma (OSCC). When patients present with a clinically positive neck, the treatment of choice is radical neck dissection. However, management of a clinically negative neck is still a subject of significant controversy.

This study was carried out in order to propose a model to predict regional lymph node metastasis of OSCC using histological parameters such as tumour stage, tumour size, pattern of invasion (POI), differentiation of tumour, and host immune response, together with the expression levels of six biomarkers (periostin, HIF-1α, MMP-9, β-catenin, VEGF-C, and EGFR), and, furthermore, to compare the impact of all these parameters on recurrence and 3 yr and 5 yr survival rates. Materials and Method. Histological materials collected from the archives were used to evaluate histological parameters and immunohistochemical profiles. Standard methods were used for immunohistochemistry and for evaluation of results. Data related to recurrence and survival (3 and 5 years) was also recorded. Clinical data was collected from patients' records.

Male to female ratio was 3 : 1. The commonest site of OSCC was the buccal mucosa, and majority of them were T3 or T4 tumours presented at stage 4. 62.5% of the tumours were well differentiated. Three-year and 5-year survival rates were significantly associated with lymph node metastasis and recurrence. POI was significantly correlated with tumour size, stage, 3-year survival, EGFR, HIF-1α, periostin, and MMP-9 (p < 0.05). Expression of EGFR showed a direct association with metastasis (p < 0.05).

POI, level of differentiation, and expression of EGFR are independent prognostic markers for lymph node metastasis. Therefore, these parameters may help in treatment planning of a clinically negative neck.

• CD31
• CD34
• Epulis
• HNC
• Ki67
• OSCC
• PROKR1/PROKR2
• VEGF/EG-VEGF

• Grade of differentiation
• Lower lip
• Neck metastasis
• Neoangiogenesis
• Squamous cell carcinoma

• Immunohistochemistry
• Lymphangiogenesis
• Squamous cell carcinoma
• Tongue
• Young adults

• Cancer
• Genotype
• Lymphedema
• Phenotype
• Risk
• Symptoms

• Ki-67 antigen
• gelatinases
• oral squamous cell carcinoma
• vascular endothelial growth factor (VEGF)-C
• vascular endothelial growth factor (VEGF)-D

• U54 GM104942 NIGMS NIH HHS

• Lymphangiogenic factor
• Lymphatic vessel density
• Lymphatic vessel dilation
• Nodal metastasis
• Oral squamous cell carcinoma

Angiogenesis is the formation of new vessels from preexisting ones which takes place by capillary sprouting. It is seen in healing, at sites of injury and collateral circulation in ischemia. It is also seen in tumors, as once the original blood supply of a tumor is exhausted it cannot grow without further blood supply. Also as the angiogenic capacity of a tumor increases, its microvasculature, that is, microvessel density (MVD) also increases. Based on this literary evidence we carried out an immunohistochemical (IHC) study to observe the relationship between the expression of vascular endothelial growth factor (VEGF) [angiogenesis] and CD 34 [MVD] in oral squamous cell carcinoma (OSCC).

To evaluate the correlation between expression of VEGF and CD 34, the role of MVD in progression of OSCC and to compare the degree of angiogenesis in different grades of OSCC.

In this study we observed the relation between angiogenesis and MVD and the overall effect of this on oral cancer.

Thirty-three cases of OSCC were stained with hematoxylin and eosin, (H and E) to confirm the diagnosis and immunohistochemically using VEGF and CD 34 antibody. The slides were evaluated for positivity and intensity of staining.

The result was subjected to statistical analysis using analysis of variance (ANOVA) test and Fisher's exact test.

VEGF positivity as well as MVD was found to be independent of the grade of the tumor. Tumor MVD was found to be independent of expression of VEGF.

• Angiogenesis
• microvessel density
• neovascularization
• oral squamous cell carcinoma

Angiogenesis, the process that leads to the formation of new blood vessels, continues to be a topic of major scientific interest. There is an increasing hope that new discoveries will lead to newer therapies that target angiogenesis as a reliable option for disease therapy.

The objective of this study was to assess the role of vascularity, correlation of morphometric aspects of vascularity, investigate its usefulness in the histopathological classification and prognosis in normal buccal mucosa (NBM), leukoplakia and squamous cell carcinoma (SCC) of buccal mucosa.

The study sample consisted of 15 cases of NBM, 30 cases of leukoplakia, and 30 cases of SCC of buccal mucosa. The 75 archival samples were stained by hemotoxylin and eosin (H and E) and Masson's trichrome (MT). The stained sections were analyzed using image analysis software.

Statistical Package for Social Sciences (SPSS) 12.0 statistical software.

The combined mean vessel density (MVD) of all the cases in H and E was 0.1112 and for MT it was 0.2150. The difference of MVD between H and E and MT was statistically significant. The mean MVD in SCC (0.3455) for MT was higher than NBM (0.1314) and leukoplakia (0.1263). The mean MVD increased from stage III (0.3563) to IV (0.5312). It also increased from NBM (0.1314) to hyperkeratosis (0.1505) and decreased from grade I (0.3556) to II (0.2795) of oral SCC (OSCC).

MVD can be used as an adjunct with other diagnostic modalities. Further studies are needed to standardize baseline levels for different sites and age groups.

• Leukoplakia
• mean vessel density
• neovascularization
• squamous cell carcinoma

Hypoxia-induced factor-1 alpha (HIF-1α) affects many effector molecules and regulates tumor lymphangiogenesis and angiogenesis during hypoxia. The aim of this study was to investigate the role of HIF-1α in the regulation of vascular endothelial growth factor C (VEGF-C) expression and its effect on lymphangiogenesis and angiogenesis in breast cancer. Lymphatic vessel density (LVD), microvessel density (MVD) and the expressions of HIF-1α and VEGF-C proteins were evaluated by immunohistochemistry in 75 breast cancer samples. There was a significant correlation between HIF-1α and VEGF-C (P = 0.014, r = 0.273, Spearman's coefficient of correlation). HIF-1α and VEGF-C overexpression was significantly correlated with higher LVD (P = 0.003 and P = 0.017, respectively), regional lymph nodal involvement (P = 0.002 and P = 0.004, respectively) and advanced tumor, node, metastasis (TNM) classification (P = 0.001 and P = 0.01, respectively). Higher MVD was observed in the group expressing higher levels of HIF-1α and VEGF-C (P = 0.033 and P = 0.037, respectively). Univariate analysis showed shorter survival time in patients expressing higher levels of HIF-1α and VEGF-C. HIF-1α was also found to be an independent prognostic factor of overall survival in multivariate analysis. The results suggest that HIF-1α may affect VEGF-C expression, thus acting as a crucial regulator of lymphangiogenesis and angiogenesis in breast cancer. This study highlights promising potential of HIF-1α as a therapeutic target against tumor lymph node metastasis.

• HIF-1α
• VEGF-C
• angiogenesis
• breast cancer
• lymphangiogenesis

The aim of the present study was to investigate the association between the expression of chemokine receptors CCR7 and CXCR4 and vascular endothelial growth factor (VEGF)-C and the lymph node metastasis of pancreatic ductal adenocarcinoma (PDAC). The mRNA transcription levels of CCR7, CXCR4 and VEGF-C were measured in 24 specimens by real-time reverse transcription (RT)-PCR, while the protein expression levels were measured in 65 specimens by immuohistochemistry. Professional software for pathological image manipulation (Image Pro Plus 6.0) was used to quantitate the results of the immunohistochemical staining. The mRNA and protein expression levels of CCR7, CXCR4 and VEGF-C were all significantly higher in the cancer samples compared with those in the adjacent normal tissue. The CCR7 and VEGF-C mRNA and protein expression levels were significantly higher in the patients with cancer types exhibiting lymph node metastasis and an advanced International Union Against Cancer (UICC) stage (P<0.05). The greater the number of metastatic lymph nodes, the higher the levels of CCR7 expression (P<0.05). There was a significant positive linear correlation between the mRNA and protein expression levels of CCR7 and VEGF-C (P<0.05). The mRNA and protein expression levels of CXCR4 were not correlated with the lymph node metastasis (P>0.05), however the strong positive expression of CCR7 and VEGF-C was significantly associated with the lymph node metastasis of PDAC.

• CCR7
• CXCR4
• VEGF-C
• lymph node metastasis
• pancreatic ductal adenocarcinoma

Head and neck squamous cell carcinoma (HNSCC) is a complex tissue that contains tumor cells and the surrounding stroma, which is populated by different types of mesenchymal cells and the extracellular matrix (ECM). Collectively, they are referred to as the tumor microenvironment (TME). Recent studies have shown that TME has a more profound influence on the growth and metastasis of HNSCC than was previously appreciated. Because carcinoma-associated fibroblasts (CAFs) are frequently observed in the stroma of the tumor, this review focuses on the potential role of tumor-CAFs interactions in progression of HNSCC. Tumor-CAFs crosstalk enhances the production of growth factors, cytokines, chemokines, matrix metalloproteinases (MMPs), and inflammatory mediators, which eventually facilitates tumor growth. In fact, factors and cells that do not support tumor growth are usually down regulated or mitigated in TME. Therefore TME may determine the fate of the tumors at the site of invasion and metastasis. For tumor cells that survive at these sites, stromal activation may serve to establish a supportive tumor stroma, fostering the outgrowth of the metastatic cells. The concept of tumor-stromal interactions and microenvironmental niche has profound consequences in tumor growth and metastasis and therefore, it's understanding will open up new strategies for the diagnosis, prognosis and therapy of HNSCC.

• CCL12.
• CXCR4
• Cancer associated fibroblasts (CAFs)
• Cycloxygenase-2 (COX-2)
• Head and neck cancer
• Matrix metalloproteinases (MMPs)

Tumors require blood supply for their growth and dissemination. It is a well accepted paradigm that tumors recruit new blood vessels from the existing circulation (angiogenesis) and this participates in tumor invasion and metastasis. Studies in the literature provide evidence for expression of Vascular Endothelial Growth Factor (VEGF) by the tumor for neo-angiogenesis, which is not only required for the tumor growth but also its metastasis. Based on the literary evidences we carried out an Immuno-Histochemical (IHC) study for VEGF in Oral Squamous Cell Carcinoma (OSCC) tissues to provide a strong link between the factor and oral cancer.

To analyze the expression of VEGF in OSCC tissues of different histological grades, clinical sizes and lymph node status and to use this as an indicator for disease progression by helping in delineating a risk population, that may benefit from an attractive adjuvant therapeutic strategy for OSCC.

Studies published from 1990 till 2010 have only seen the association of VEGF with tumor angiogenesis and its possible role in metastasis. This is the first study that takes into account the clinical status of the lymph nodes and VEGF expressivity in a sample size of 30 cases.

30 oral squamous cell carcinoma tissue slides were stained using Hematoxylin and Eosin stain (to confirm the diagnosis) and immunohistochemically using VEGF antibody. IHC stained slides were thereafter evaluated for the positivity and intensity.

The result was subjected to statistical analysis using Chi-square test

VEGF positivity was seen in approximately. 90% of cases which was independent of histological grade of OSCC. However the intensity increased with the clinical size of cancer and from palpable lymph node to a tender and hard lymph node.

• Angiogenesis
• VEGF
• immunohistochemistry

The prediction of tumor behavior for patients with oral carcinomas remains a challenge for clinicians. The presence of lymph node metastasis is the most important prognostic factor but it is limited in predicting local relapse or survival. This highlights the need for identifying biomarkers that may effectively contribute to prediction of recurrence and tumor spread. In this study, we used one- and two-dimensional gel electrophoresis, mass spectrometry and immunodetection methods to analyze protein expression in oral squamous cell carcinomas. Using a refinement for classifying oral carcinomas in regard to prognosis, we analyzed small but lymph node metastasis-positive versus large, lymph node metastasis-negative tumors in order to contribute to the molecular characterization of subgroups with risk of dissemination. Specific protein patterns favoring metastasis were observed in the “more-aggressive” group defined by the present study. This group displayed upregulation of proteins involved in migration, adhesion, angiogenesis, cell cycle regulation, anti-apoptosis and epithelial to mesenchymal transition, whereas the “less-aggressive” group was engaged in keratinocyte differentiation, epidermis development, inflammation and immune response. Besides the identification of several proteins not yet described as deregulated in oral carcinomas, the present study demonstrated for the first time the role of cofilin-1 in modulating cell invasion in oral carcinomas.

• Aged
• Carcinoma, Squamous Cell/metabolism
• Carcinoma, Squamous Cell/pathology
• Cofilin 1/genetics
• Cofilin 1/metabolism
• Electrophoresis, Gel, Two-Dimensional
• Electrophoresis, Polyacrylamide Gel
• Female
• Gene Knockdown Techniques
• Humans
• Immunohistochemistry
• Lymphatic Metastasis
• Male
• Mass Spectrometry
• Middle Aged
• Mouth Neoplasms/metabolism
• Mouth Neoplasms/pathology
• Neoplasm Invasiveness
• Proteomics

• oral cancer
• gene expression profiling
• lymph node metastasis
• microarray
• diagnosis and prognosis
• biomarker

• R03 CA135992-02 NCI NIH HHS
• R01 CA139596-03 NCI NIH HHS
• R03 CA114688 NCI NIH HHS
• R01 CA139596 NCI NIH HHS
• K22 DE014847-05 NIDCR NIH HHS
• K22 DE014847 NIDCR NIH HHS
• R03 DE016569-03 NIDCR NIH HHS
• R03 CA135992 NCI NIH HHS
• R03 DE016569 NIDCR NIH HHS
• R03 CA114688-03 NCI NIH HHS

• Carcinoma, Squamous Cell
• Humans
• Leukoplakia, Oral/complications
• Leukoplakia, Oral/diagnosis
• Leukoplakia, Oral/therapy
• Mouth Diseases/diagnosis
• Mouth Diseases/etiology
• Mouth Diseases/therapy
• Mouth Mucosa
• Skin Diseases/diagnosis
• Skin Diseases/etiology
• Skin Diseases/therapy

• Aged
• Biomarkers, Tumor/analysis
• Biomarkers, Tumor/metabolism
• Carcinoma, Squamous Cell/blood supply
• Carcinoma, Squamous Cell/metabolism
• Carcinoma, Squamous Cell/pathology
• Female
• Gene Expression Regulation, Neoplastic
• Humans
• Immunohistochemistry
• Lymphangiogenesis
• Lymphatic Metastasis
• Lymphatic Vessels/pathology
• Male
• Microcirculation
• Middle Aged
• Mouth Neoplasms/blood supply
• Mouth Neoplasms/metabolism
• Mouth Neoplasms/pathology
• Neovascularization, Pathologic/metabolism
• Predictive Value of Tests
• Prognosis
• Survival Analysis
• Time Factors
• Up-Regulation
• Vascular Endothelial Growth Factor C/analysis
• Vascular Endothelial Growth Factor C/metabolism

AIM: To clarify the usefulness of immunohistochemical molecular markers in predicting lymph node metastasis of submucosal colorectal cancer.

METHODS: We examined microvessel density, lymphatic vessel density, the Ki-67 labeling index, expression of MUC1 and Matrix metalloproteinase-7 (MMP-7) in tumor cells, and expression of cathepsin D in stromal cells at the invasive front by immunostaining of samples resected from 214 patients with submucosal colorectal cancer. Pathologic features were assessed on hematoxylin-eosin-stained samples. We evaluated the relations between clinicopathologic/immunohistochemical features and lymph node metastasis.

RESULTS: Lesions of the superficial type, with an unfavorable histologic grade, budding, lymphatic involvement, high microvessel density (≥ 40), high lymphatic vessel density (≥ 9), high Ki-67 labeling index (≥ 42), and positivity of MUC1, cathepsin D, and MMP-7 showed a significantly high incidence of lymph node metastasis. Multivariate analysis revealed that high microvessel density, unfavorable histologic grade, cathepsin D positivity, high lymphatic vessel density, superficial type, budding, and MUC1 positivity were independent risk factors for lymph node metastasis. A combined examination with four independent immunohistochemical markers (microvessel density, cathepsin D, lymphatic vessel density, and MUC1) revealed that all lesions that were negative for all markers or positive for only one marker were negative for lymph node metastasis.

CONCLUSION: Analysis of a combination of immuno-histochemical molecular markers in endoscopically resected specimens of submucosal colorectal cancer allows prediction of curability regardless of the pathologic features visible of hematoxylin-eosin-stained sections.

• Submucosal colorectal cancer
• Microvessel density
• Lymphatic vessel density
• Mucin 1
• Ki-67
• Cathepsin D
• Matrix metalloproteinase-7
• Lymph node metastasis
• Immunohistochemistry

• Biomarkers, Tumor/analysis
• Colonoscopy
• Colorectal Neoplasms/pathology
• Colorectal Neoplasms/surgery
• Humans
• Immunohistochemistry
• Logistic Models
• Lymphatic Metastasis/pathology
• Prognosis

• Adult
• Aged
• Aged, 80 and over
• Antibodies, Monoclonal
• Antibodies, Monoclonal, Murine-Derived
• Antigens, CD34
• Carcinoma, Squamous Cell/blood supply
• Carcinoma, Squamous Cell/chemistry
• Carcinoma, Squamous Cell/pathology
• Female
• Humans
• Lymphangiogenesis/physiology
• Lymphatic Vessels/chemistry
• Lymphatic Vessels/pathology
• Male
• Membrane Glycoproteins
• Middle Aged
• Mouth Neoplasms/blood supply
• Mouth Neoplasms/chemistry
• Mouth Neoplasms/pathology
• Neoplasm Proteins/analysis
• Neovascularization, Pathologic/pathology
• Vascular Endothelial Growth Factor A/analysis
• Vascular Endothelial Growth Factor C/analysis

• Adult
• Aged
• Aged, 80 and over
• Carcinoma, Squamous Cell/metabolism
• Carcinoma, Squamous Cell/pathology
• Female
• Humans
• Immunoenzyme Techniques
• Lymphangiogenesis/physiology
• Lymphatic Metastasis/diagnosis
• Male
• Middle Aged
• Mouth Neoplasms/metabolism
• Mouth Neoplasms/pathology
• Neoplasm Staging
• Prognosis
• ROC Curve
• Regression Analysis
• Retrospective Studies
• Sensitivity and Specificity
• Vascular Endothelial Growth Factor C/analysis
• Vascular Endothelial Growth Factor C/biosynthesis
• Vascular Endothelial Growth Factor D/analysis
• Vascular Endothelial Growth Factor D/biosynthesis
• Vascular Endothelial Growth Factor Receptor-3/analysis
• Vascular Endothelial Growth Factor Receptor-3/biosynthesis
• Vascular Endothelial Growth Factors/analysis
• Vascular Endothelial Growth Factors/biosynthesis

• Carcinoma, Squamous Cell/metabolism
• Carcinoma, Squamous Cell/mortality
• Carcinoma, Squamous Cell/radiotherapy
• Head and Neck Neoplasms/metabolism
• Head and Neck Neoplasms/mortality
• Head and Neck Neoplasms/radiotherapy
• Humans
• Kaplan-Meier Estimate
• Male
• Middle Aged
• Neoplasm Staging
• Prognosis
• Retrospective Studies
• Survival Analysis
• Vascular Endothelial Growth Factor D/metabolism
• Vascular Endothelial Growth Factor D/radiation effects
• Vascular Endothelial Growth Factors/metabolism
• Vascular Endothelial Growth Factors/radiation effects