Immune-phenotyping in Hypoxic Microenvironment: A Prelude to Oral Carcinogenesis

The purpose of this study was to investigate the association of CD8⁺ tumor-infiltrating lymphocytes (CD8⁺ TILs), microvascular density (MVD), and vascular endothelial growth factor (VEGF) with the TNM (Tumor-Node-Metastasis) stage, as well as to analyze their interrelationships in colorectal adenocarcinoma. This cross-sectional study involved 50 FFPE samples of colorectal adenocarcinoma from the Laboratory of Anatomical Pathology at Dr. Soedarso Hospital. Microvascular density (MVD) was assessed on hematoxylin-eosin (H&E) slides, while CD8⁺ tumor-infiltrating lymphocytes (CD8⁺ TILs) and vascular endothelial growth factor (VEGF) expression were evaluated using immunohistochemistry (IHC). Correlation analysis was conducted between the biomarkers and the TNM stage, including its components depth of tumor invasion, lymph node status, and distant metastasis using the Chi-square test. Spearman's correlation was used to assess the relationships among CD8⁺ TILs, MVD, and VEGF. High CD8⁺ TILs expression was significantly associated with negative lymph node status (p = 0.047; OR = 0.31, 95% CI = 0.098-1.001), absence of distant metastasis (p = 0.008; OR = 0.130, 95% CI = 0.025-0.680), and low TNM stage (p = 0.011; OR = 0.221, 95% CI = 0.067-0.727). The distribution of high MVD was correlated with deeper tumor invasion (p = 0.004; OR = 9.036, 95% CI = 1.741-46.890), positive lymph node status (p < 0.001; OR = 10.286, 95% CI = 2.768-38.215), and high TNM stage (p = 0.002; OR = 6.612, 95% CI = 1.924-22.728). High expression of VEGF showed a significant correlation with deeper tumor invasion (p = 0.036; OR = 0.675, 95% CI = 0.544-0.837). Spearman's test revealed a negative correlation between CD8⁺ TILs and MVD (r = -0.280, p = 0.049), and a positive correlation between MVD and VEGF (r = 0.303, p = 0.032). High CD8⁺ TILs and low MVD are favorable prognostic factors in colorectal adenocarcinoma, whereas increased MVD and VEGF expression indicate tumor aggressiveness and enhanced angiogenesis. The combination of immune and angiogenic biomarkers with TNM staging could improve prognostic evaluation in colorectal adenocarcinoma.

Oral squamous cell carcinoma (OSCC) is one kind of invasive malignancy arising from oral epithelial cells. According to the statistical data of the Department of Health in Taiwan, OSCC is most common cancer in head and neck and also the fifth leading cause of cancer mortality in 2010. In order to promote their own growth and dispersion, malignant tumors will secrete variant signaling molecules to recruit host blood vessels to grow into the vicinity of the tumor (so-called tumor angiogenesis). Among these signaling molecules, vascular endothelial growth factor (VEGF) is the most important and usually overexpresses in head and neck squamous cell carcinoma. In OSCC, VEGF highly correlates with microvessel density (MVD) of oral cancer micro-environment, and negatively correlates with the survival rates of OSCC patients. Besides, endothelin-1 (ET-1) composed of 21 amino acids plays an important role in tumor growth, metastases, and angiogenesis. Furthermore, it is proved that ET-1 released by tumors would activate endothelin A receptor (ETAR) of the tumor cells and induce VEGF expression in ovarian. Moreover, studies indicated that the expression of ET-1 is increased in OSCC cell lines. However, the relationship between ET-1 and VEGF in OSCC is still not clear. Therefore, in this study we want to testify that whether ET-1 will modulate the expression of VEGF in OSCC via ETAR, and also confirm this phenomenon in clinical samples at the same time. Firstly, we find the significant correlation between ET-1 and VEGF expressions with immunohistochemical stain in clinical OSCC samples. In survival analysis, the patients with high VEGF expressions, instead of ET-1, have lower survival rates, and there is a statistical significance. In the studies of OSCC cell lines, after ET-1 stimulated, the expression of VEGF was increased in OSCC cells, and was inhibited by ETAR antagonist (BQ-123). This phenomenon indicated that ET-1 would induce the expression of VEGF via ETAR in OSCC, and the basal expressions of ET-1 and VEGF would regulate this result. On the other hand, VEGF expression did not inhibit completely by ETAR antagonist when compared with the control group or the group with BQ-123 only. We speculated that ET-1 might be a strong agonist, or the other receptor of ET-1, endothelin B receptor (ETBR), had the similar function as ETAR did. However, the expression of VEGF is regulated by many factors, so therefore we need to study the underlying mechanism of carcinogenesis. Finally, the basal expressions of mRNA of ET-1 and VEGF were varied between selected nine OSCC cell lines. There was a positive correlation between ET-1 and VEGF when we omitted OEC-M1 and OC-2 which expressed great levels of VEGF mRNA. This result matched the finding of clinical samples. According to the experimental results, ETAR antagonist will inhibit the expression of VEGF in OSCC; however, it still further studies whether ETAR antagonist can become a candidate of the anti-cancer drug.

AIMS: To correlate vascular endothelial growth factor (VEGF) expression in oral squamous cell carcinoma with the clinicopathological characteristics and prognosis; and to assess whether p53 gene status is associated with...

Cellular senescence prevents the proliferation of cells at risk for neoplastic transformation. Nonetheless, the senescence response is thought to be antagonistically pleiotropic and thus contribute to aging phenotypes, including, ironically, late life cancers. The cancer-promoting activity of senescent cells is likely due to secreted molecules, the identity of which remains largely unknown. Here, we have shown that senescent fibroblasts, much more than presenescent fibroblasts, stimulate tumor vascularization in mice. Weakly malignant epithelial cells co-injected with senescent fibroblasts had larger and greater numbers of blood vessels compared with controls. Accordingly, increased vascular endothelial growth factor (VEGF) expression was a frequent characteristic of senescent human and mouse fibroblasts in culture. Importantly, conditioned medium from senescent fibroblasts, more than medium from presenescent cells, stimulates cultured human umbilical vein endothelial cells to invade a basement membrane, a hallmark of angiogenesis. Increased VEGF expression was specific to the senescent phenotype and increased whether senescence was induced by replicative exhaustion, overexpression of p16(Ink4a), or overexpression of oncogenic RAS. The senescence-dependent increase in VEGF production was accompanied by very little increase in hypoxic-inducible (transcription) factor 1 alpha protein levels, and hypoxia further induced VEGF in senescent cells. This result suggests the rise in VEGF expression at senescence is not a hypoxic response. Our findings may in part explain why senescent cells stimulate tumorigenesis in vivo and support the idea that senescent cells may facilitate age-associated cancer development by secreting factors that promote malignant progression.

Vascular endothelial growth factor (VEGF) expression is associated with malignancy progression, metastasis, and poor prognosis in many malignancies, including osteosarcoma. However, studies concerning correlations between VEGF expression and histopathological prognostic factors ofosteosarcoma are limited. This study aimed to evaluate the correlations between VEGF expression and histopathological findings in osteosarcoma’spatients.This was a cross-sectional study using formalin-fixed paraffin embedded (FFPE) samples of 32 osteosarcoma’s patients from Dr. Sardjito General Hospital, Yogyakarta. Histopathological findings of specimens were re-evaluated by two independent observers, recorded for the subtypes, invasiveness, grading, mitotic counts, and tumor infiltrating lymphocytes (TIL). Expression of VEGF was determined based on immunostaining and evaluated using immunoreactivity score (IRS).Chi-square and Spearman correlation test were used to analyze the association between variables. Range of VEGF expression score was 0 to 11, with mean 5.09. Significant negative correlation between the VEGF expression and TIL was observed (p=0.046). However, there was no significant correlations between the VEGF expression and osteosarcomas subtypes, invasion, grading or mitotic counts (p&gt; 0.05). In conclusion, the VEGF expression is associated with TIL. Further study is needed to evaluate the roles of VEGF and lymphocytes in osteosarcoma development dan progression in order to better understand of the role of VEGF in immunotherapy of osteosarcoma.

Vascular endothelial growth factor (VEGF) is a potent mitogen and permeability factor for endothelial cells that plays a central role in angiogenesis, vascular maintenance, inflammation, and cancer. VEGF also mediates the homeostatic adaptation to hypoxic conditions by promoting an increase in vascular density to compensate for decreased oxygenation. This process is triggered by an oxygen-sensitive transcription factor, hypoxia-inducible factor-1 (HIF1alpha), which becomes active in hypoxic tissues, leading to the synthesis and secretion of VEGF. The role of HIF1alpha in other processes that involve angiogenesis such as in inflammation is less clear. Of interest, endothelial cells not only respond to but also store and secrete VEGF, which is required for the maintenance of the integrity of the vascular system. How this intracellular pool of VEGF is regulated is still not understood. Here, we found that CXCL8/IL8, a potent proangiogenic and inflammatory chemokine, up-regulates VEGF mRNA and protein levels in endothelial cells by acting on its cognate receptor, CXCR2, and that this results in the autocrine activation of VEGFR2. Surprisingly, this process does not involve HIF1alpha but instead requires the activation of the transcription factor NFkappaB. Furthermore, we identified the components of the CBM complex, Carma3, Bcl10, and Malt1, as key mediators of the CXCL8/IL8-induced NFkappaB activation and VEGF up-regulation. Together, these findings support the existence of an NFkappaB-mediated pathway by which the proinflammatory chemokine CXCL8/IL8 controls the expression of VEGF in endothelial cells, thereby promoting the activation of VEGF receptors in an autocrine fashion.

Correlations of TNM staging and lymph node metastasis of gastric cancer with MRI features and VEGF expression

Prognostic Value of the VHL, HIF-1α, and VEGF Signaling Pathway and Associated MAPK (ERK1/2 and ERK5) Pathways in Clear-Cell Renal Cell Carcinoma. A Long-Term Study.

Etiology of OHSS and use of dopamine agonists

Breast cancer is the most frequent malignancy in women worldwide. In spite of its questionable reliability, the TNM (Tumor, Node, Metastasis) staging system is widely used for the prognosis of this disease. On the other hand, angiogenesis is considered to have an essential role in the evolution of breast cancer and the Vascular Endothelial Growth Factor (VEGF) has proven to be the key regulator of this process. Quantitation of VEGF and the tumor vasculature might play an important role in predicting tumor behavior and patient management. The aim of our study was to evaluate the potential correlation between the VEGF expression, microvessel density (MVD) in the tumor tissues and TNM staging in breast cancer. We included 34 patients with breast cancer who had undergone surgical treatment and evaluated each case clinically, histopathologically and by immunohistochemistry for VEGF and CD31 expression in the tumor tissues. VEGF expression was evaluated by calculating the average percentage of cytoplasmic positive cells from 3 high power fields. MVD was expressed as the average number of the CD31+ microvessels from 3 high power fields. Expression of VEGF was significantly associated with the number of lymph nodes with metastasis, the number of cells in mitosis, the presence of necrosis and the T-stage (inverse correlation). MVD correlated very well with the histological grading, the number of cells in mitosis, the presence of inflammation and the presence of necrosis. No correlation could be established between VEGF expression and MVD. Although their relationship with TNM staging remains unclear, VEGF expression and MVD proved to be important indicators of the malignant status in breast cancer, confirming the major involvement of angiogenesis in this type of cancer. Both of them are valuable prognostic factors in breast cancer, but the pattern of their relationship needs further analysis of the VEGF receptors in order to be described.

To investigate the clinicopathological role of expression of vascular endothelial growth factor (VEGF) and cortactin, as well as whether their expression are independent predictors of tumor recurrence following curative resection of gastric cancer. One hundred twenty-eight patients with gastric cancer were included in this study. Formalin-fixed paraffin-embedded specimens were stained for VEGF and cortactin, and the correlation between the staining, clinicopathological parameters and prognostic power were analyzed. Of the 128 patients studied, 58 (45.3%) and 71 (55.5%) cases were strongly positive for VEGF and cortactin, respectively. VEGF expression correlated with Lauren classification (P < 0.001), pathological tumor stage (P < 0.001), and pathological tumor node metastasis (TNM) stage (P = 0.003). Cortactin expression correlated with pathological lymph node stage (P = 0.018), pathological TNM stage (P < 0.001), and degree of differentiation (P < 0.001). There were statistically significant associations between tumor recurrence and VEGF expression (P = 0.023), and cortactin expression (P < 0.001). In multivariate analysis, pathological TNM stage, VEGF expression, and cortactin expression were independent prognostic influence on disease-free survival (P < 0.001, 0.022, and 0.034, respectively). VEGF and cortactin may be a good biomarker to be applied in clinic to predict the prognosis of patients with curatively resected gastric cancer.

Background: Overexpression of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) and increasing mast cell density (MCD) in premalignant and malignant oral lesions have been documented. However, their correlation with clinicopathologic parameters and survival rate in oral squamous cell carcinoma (OSCC) is not completely clear. This study aimed to assess these subjects. Methods: VEGF, COX-2, and mast cell tryptase expression were examined immunohistochemically in 57 cases of OSCC. The relationships between the markers' expression and clinicopathologic data were assessed using bivariate and multivariate analysis. Results: Spearman's rank correlation coefficient showed a significant correlation between VEGF and COX-2 expression (r = 0.462, p < 0.001), as well as between VEGF expression and MCD (r = 0.306, p < 0.001). Multivariate analysis showed no significant correlation between the markers' immunoexpression and overall survival (OS), but a significant correlation between mode of invasion and OS [hazard ratio 0.362 (95% CI: 0.138- 0.974); p = 0.038] was observed. An association between MCD and gender (p = 0.042) was also found, as MCD was higher in males. Conclusion: The significant correlation of VEGF expression with COX-2 expression and MCD may represent the roles of COX-2 and MCD in tumor angiogenesis by modulating VEGF production. However, VEGF, COX-2, and MCD are not useful indicators to predict prognosis in OSCC. Nevertheless, the mode of invasion can be considered as an independent prognostic factor in OSCC patients.

Correlation of hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) expressions with clinico-pathological features of oral squamous cell carcinoma (OSCC)

Predictive factor for photodynamic therapy effects on oral squamous cell carcinoma and oral epithelial dysplasia

Tumor-infiltrating lymphocytes in oral cavity squamous cell carcinoma and its association with clinicopathological parameters