Are basic fibroblast growth factor and vascular endothelial growth factor prognostic indicators in pediatric patients with malignant solid tumors?

Angiogenesis plays an important role in the growth, progression, and metastasis of solid tumors. Among angiogenic factors, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) appear to be useful markers in adults with cancer. The aim of this pilot study was to determine the levels of VEGF in serum and bFGF in serum and urine of children with solid tumor at diagnosis (as measured by ELISA), and to investigate whether these parameters provide prognostic information. Forty consecutive patients with different types of cancer were prospectively included in this study. Median values of all studied angiogenic factors were higher in patients than in controls (n = 40), and the differences were statistically significant for bFGF in serum and urine: 10 versus 3 pg/ml (P = 0.0004) and 6406 versus 0 pg/g of creatinine (P < 0.0001), respectively. Among patients, median serum values of bFGF and VEGF were higher in children with metastatic disease (n = 14) than in those with localized disease (n = 26). The difference was statistically significant for serum bFGF: 17.5 versus 6 pg/ml (P = 0.02). Serum angiogenic factor levels correlated with outcome. The estimated event-free survival at 3 years was 79% for patients with normal bFGF values (n = 13) versus 42% (n = 26; P = 0.02) for those with high levels, and 71% in case of normal VEGF values (n = 20) versus 38% (n = 19; P = 0.04) for those with high levels. No benefit of normal urinary bFGF values was observed. Our results provide a rationale for exploring the clinical interest of bFGF and VEGF measurements in body fluids of a larger group of children with cancer.     

Polymorphism of vascular endothelial growth factor �C2578C/A with cancer risk: evidence from 11263 subjects

Published data on the association between vascular endothelial growth factor (VEGF) ?C2578C/A polymorphism and cancer risk is inconclusive. To derive a more precise estimation of association between VEGF ?C2578C/A polymorphism and the risk of cancer, we performed a meta-analysis of 5415 cancer cases and 5848 controls from 16 published case?Ccontrol studies. We used odds ratios (ORs) with 95% confidence intervals (CIs) to assess the strength of the association. Our meta-analysis indicated that VEGF ?C2578C/A polymorphism was associated with the risk of colorectal cancer under homozygote comparison (OR?=?0.70, 95% CI?=?0.53?C0.92), dominant model (OR?=?0.72, 95% CI?=?0.57?C0.92), and recessive model (OR?=?0.82, 95% CI?=?0.67?C1.01), although no evidence of association between VEGF ?C2578C/A polymorphism and cancer risk was observed as we compared in the pooled analyses (homozygote comparison: OR?=?0.97, 95% CI?=?0.81?C1.16). More studies are needed to detect VEGF ?C2578C/A polymorphism and its association with cancer in different ethnic populations incorporated with environmental exposures in the susceptibility of different kinds of cancer.     

Basic fibroblast growth factor and vascular endothelial growth factor serum levels in breast cancer patients and healthy women: useful as diagnostic tools?

Introduction  

The aim of the present study was to analyze the relationship between the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in breast cancer cells and the corresponding serum levels in individual patients. The study also evaluated the potential of serum levels of the two growth factors as diagnostic markers in a case–control study.     

Clinical significance of vascular endothelial growth factor‐C (VEGF‐C) in breast cancer

Vascular endothelial growth factorC (VEGFC) is a specific ligand which induces lymphangiogenesis. We examined the expression of VEGFC protein to determine its role in the progression of breast cancer. Immunohistochemical analysis revealed that VEGFC was overexpressed in 39 of 98 breast cancer specimens (39.8%) but not in adjacent normal mammary glands. The expression of VEGFC showed a significant correlation with lymphatic vessel invasion (p=0.0004). It is noteworthy that the 5year disease free survival rate of the VEGFC positive group was significantly poorer than that of negative group (p=0.0356). We suggest that as expression of VEGFC is not implicated in lymphatic spread, it may prove to be a promising marker to predict the recurrence of breast cancer.     

Vascular endothelial growth factor +936C/T polymorphism and breast cancer risk: a meta-analysis of 13 case–control studies

The association between vascular endothelial growth factor (VEGF) +936C/T polymorphism and breast cancer risk has been widely reported, but results were inconsistent. In order to derive a more precise estimation of the relationship, a meta-analysis was performed. Eligible articles were identified through search of databases including PubMed, Embase, and Chinese Biomedical Literature Database (CBM). The association between the VEGF +936C/T polymorphism and breast cancer risk was conducted by odds ratios (ORs) and 95 % confidence intervals (95 % CIs). Finally, a total of 13 studies with 6,879 cases and 7,219 controls were included in our meta-analysis. Overall, a significant association was found between VEGF +936C/T polymorphisms and the risk of breast cancer in overall populations under five models (T vs. C: OR?=?0.83, 95 % CI?=?0.73–0.94, P?=?0.002; TT vs. CC: OR?=?0.74, 95 % CI?=?0.61–0.91, P?=?0.004, Fig. 1a; TC vs. CC: OR?=?0.83, 95 % CI?=?0.71–0.96, P?=?0.014; TT vs. CC/CT: OR?=?0.77, 95 % CI?=?0.62–0.94, P?=?0.010; TT/TC vs. CC: OR?=?0.82, 95 % CI?=?0.72–0.95, P?=?0.006). In the subgroup analysis by ethnicity, there were also significant associations found between VEGF +936C/T polymorphism and breast cancer risk in Asians and Caucasians. In conclusion, the results of our meta-analysis suggest that the VEGF +936C/T polymorphism is significantly associated with breast cancer development and the VEGF 936T allele carriers may be associated with decreased breast cancer risk.     

Interleukin 8 and vascular endothelial growth factor -- prognostic factors in human gastric carcinomas?

Gastric carcinoma cells express potent angiogenic factors including vascular endothelial growth factor (VEGF). We previously reported that interleukin-8 (IL-8) acts as an angiogenic factor for human gastric carcinomas. More recently, we found that IL-8 upregulates matrix metalloproteinase-9 (MMP-9) expression and increases invasive activity of gastric carcinoma cells. The purpose of this study was to determine whether the expression of IL-8 and VEGF correlates with clinicopathological parameters in human gastric carcinomas. IL-8 and VEGF expression levels were measured by an enzyme-linked immunosorbent assay (ELISA) in 56 gastric carcinomas and the surrounding normal mucosa. Macroscopic and histopathological tumour findings, presence of metastasis and prognosis were obtained from the patient records and endoscopic, surgical and pathological reports. IL-8 protein levels were higher in most neoplasms than in the corresponding normal mucosal tissue. In contrast, VEGF expression in the tumours was similar to that in normal mucosa. The IL-8 level in the neoplasms correlated significantly with the depth of invasion, venous invasion and lymphatic invasion. VEGF expression in the tumours correlated well with the depth of invasion and lymph node metastasis. No correlation between IL-8 and VEGF expression in the tumours was observed. The survival rates of patients with tumours displaying high IL-8 and VEGF expression levels were significantly lower (P<0.05) than those of patients with tumours displaying low IL-8 and VEGF expression. The results suggest that IL-8 and VEGF may be independent and important prognostic factors in human gastric carcinomas.     

Stromal interaction essential for vascular endothelial growth factor A-induced tumour growth via transforming growth factor-β signalling

Background:

High vascular endothelial growth factor (VEGFA) levels at the time of diagnosis confer a worse prognosis to multiple malignancies. Our aim was to investigate the role of VEGFA in promoting tumour growth through interaction with its environment.

Methods:

HL-60 cells were transduced with VEGFA165 or control vector using retroviral constructs. Control cells (n=7) or VEGFA165 cells (n=7) were subcutaneously injected into NOD/SCID mice. Immunohistochemistry of markers for angiogenesis (CD31) and cell proliferation (Ki67) and gene expression profiling of tumours were performed. Paracrine effects were investigated by mouse-specific cytokine arrays.

Results:

In vivo we observed a twofold increase in tumour weight when VEGFA165 was overexpressed (P=0.001), combined with increased angiogenesis (P=0.002) and enhanced tumour cell proliferation (P=0.001). Gene expression profiling revealed human genes involved in TGF-β signalling differentially expressed between both tumour groups, that is, TGFBR2 and SMAD5 were lower expressed whereas the inhibitory SMAD7 was higher expressed with VEGFA165. An increased expression of mouse-derived cytokines IFNG and interleukin 7 was found in VEGFA165 tumours, both described to induce SMAD7 expression.

Conclusion:

These results suggest a role for VEGFA-driven tumour growth by TGF-β signalling inhibition via paracrine mechanisms in vivo, and underscore the importance of stromal interaction in the VEGFA-induced phenotype.     

Effect of vascular endothelial growth factor on remodeling of C6 glioma tissue in vivo

Pathological characteristics and biological behaviors of tumor are considered to result from pathological tissue remodeling regulated by the interaction of factors in the tumor microecosystem (TMES). Vascular endothelial growth factor (VEGF) is one of the factors that probably play an important role in the process of tissue remodeling. This study was a comprehensive investigation of the effects of VEGF on remodeling of glioma tissue in vivo. C6 cells with expression vectors containing sense (C6/VEGF+) or antisense (C6/VEGF−) VEGF₁₆₄ complementary DNA (cDNA) or an empty vector (C6/vec) were implanted into athymic mice, which served as an in vivo model with different levels of VEGF expression. VEGF expression, water content, and morphological characteristics of these tumor tissues were assayed. Expression of VEGF and water content in C6/VEGF− glioma (C6/VEGF−G) tissues were lower than in C6, C6/VEGF+, and C6/vec glioma (C6G, C6/VEGF+G, C6/vecG) (P < 0.01, P < 0.05); water content correlated with VEGF expression (r = 0.791, P = 0.000). In C6/VEGF−G, tumor cells were not tightly adhered to vascular walls, and the basal lamina surrounded by collagen fibers was monolayer and not continuous. However, in C6G, C6/VEGF+G, and C6/vecG, tumor cells were very close to the vascular walls, with some extending their processes to the wall. Generally, loose basal laminae surrounded by small amounts of collagen fibers were multilayer, integrated, and continuous. Vesicular vacuolar organelle (VVO) structures were observed in plasma of vascular endothelial cells (VECs), and significant correlation was found between VEGF expression level and VVO density (Spearman’s r = 0.642, P = 0.007). No correlation was found between VEGF expression and fenestrae formation in VECs (Pearson’s correlation r = −0.053, P = 0.846), and fenestrae in neither VECs nor intercellular clefts correlated with water content of tumor tissue (Pearson’s correlation r = 0.018, P = 0.947). These results demonstrate that VEGF can aggravate edema in tumor tissues by increasing VVOs and plays critical roles in the stickiness of tumor cells to vessel wall and in the integrity and continuity of the basal lamina of vessels. Our data indicate a possible mechanism of remodeling of glioma tissue and suggest that blocking VEGF might contribute to a therapeutic strategy for glioma.     

Hypoxia-inducible factor-1α and vascular endothelial growth factor expression in circulating tumor cells of breast cancer patients

Introduction  

The detection of peripheral blood circulating tumor cells (CTCs) and bone marrow disseminated tumor cells (DTCs) in breast cancer patients is associated with a high incidence of disease relapse and disease-related death. Since hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) play an important role in angiogenesis and tumor progression, the purpose of the current study was to investigate their expression in CTCs.     

Does vascular endothelial growth factor (VEGF) predict local relapse and survival in radiotherapy-treated node-negative breast cancer?

The aim of this study was to determine the association of vascular endothelial growth factor (VEGF) content in 302 consecutive node-negative breast cancer (NNBC) patients treated with only locoregional radiotherapy to relapse free- (RFS) and overall survival (OS). VEGF content in tumour cytosols was measured by an enzymatic immunoassay for the major isoform VEGF165. The median age was 56 years, the median follow-up time 56 months. A wide range (0.01-144.79 pg microg(-1) DNA) of VEGF content was found (median 1.92). Significant associations were found between VEGF and oestrogen receptor (ER) content, progesterone receptor (PR) and tumour size (P = 0.005). Univariate analysis displayed significant reduced RFS and OS for patients with higher VEGF content (P = 0.0113 and P = 0.0075 respectively). A total of 43 recurrences have been found (ten local relapses within the breast, five in the axillary or supraclavicular lymph nodes and 28 distant metastasis). There was no significant correlation between the localization of the relapse and the VEGF content. Multivariate analysis suggested VEGF as the only predictor of OS (relative risk (RR) = 3.6, 95% confidence interval (CI) = 0.97-13.37), and in patients with T1 tumours (n = 236) the multivariate analysis clearly displayed VEGF as the only independent predictor of both RFS and OS (RR = 5.1, CI = 1.07-24.59). In the subgroup with ER-positive tumours (n = 229), multivariate analysis showed VEGF as the only significant predictor of RFS and OS (RR = 10.44, CI = 1.26-86.38). The results suggest VEGF165 as a predictor of RFS and OS in NNBC patients treated with locoregional radiotherapy, comprising especially patients with favourable prognosis of T1 tumours, or ER-positive tumours. The high VEGF expression might define a radioresistant phenotype, or indicate an early distant spread which might require adjuvant systemic treatment.     

Vascular endothelial growth factor receptors 1,3 and caveolin-1 are implicated in colorectal cancer aggressiveness and prognosis—correlations with epidermal growth factor receptor, CD44v6, focal adhesion kinase, and c-Met

Vascular endothelial growth factor receptor-1 (VEGFR-1) and caveolin-1 have been shown to act both as tumor-promoting and tumor-suppressing proteins in various malignancies as well as in colorectal cancer (CRC), while VEGFR-3’s lymphagiogenic involvement and connection to tumor parameters has yielded heterogenic results. This study was designed to investigate the expression of these molecules in 183 human CRC tissue specimens and explore their effect in both clinicopathological parameters and disease prognosis. We also utilize our previous results regarding epidermal growth factor receptor (EGFR), c-Met, CD44v6, and focal adhesion kinase, in an attempt to further clarify their distinct role in tumor prognosis and their crosstalk. Caveolin-1 was more freely distributed in the neoplasms of the right colon and restricted towards the left and the rectal cancer samples (p?=?0.022); VEGFR-3 was associated with higher nodal metastasis’ status (p?=?0.001) and staging (p?=?0.006), and loss of VEGFR-1 predicted distant metastasis (p?=?0.026) and advanced stage (p?=?0.049). Prompted by previous reports, we performed all analyses also in the patient group of early (I and II) tumor stage where it was evident that VEGFR-1 was more frequently expressed in patients under 60 years old (p?=?0.014) and VEGFR-3 was significantly elevated in left colon cancers (p?=?0.039) and female patients (p?=?0.038). Within the advanced stage (III and IV), the absence of VEGFR-1 exhibited a tendency for higher M status (p?=?0.067) and lack of caveolin-1 signified worse AJCC classification (p?=?0.053). Additionally, patient survival was influenced from VEGFR-3 (p?=?0.019) for the whole sample, whereas subgroup analyses provided a correlation between caveolin-1 expression and improved survival in the early detection group of patients (p?=?0.022). Using Cox regression for all available markers, EGFR, CD44v6, and VEGFR-1 emerged in this study as potential surrogate markers, the latter having positive prognostic significance. We further explored the multiple receptor correlations that were identified.     

CdGAP is required for transforming growth factor β- and Neu/ErbB-2-induced breast cancer cell motility and invasion

RhoA, Rac1 and Cdc42, the best-characterized members of the Rho family of small GTPases, are critical regulators of many cellular activities. Cdc42 GTPase-activating protein (CdGAP) is a serine- and proline-rich RhoGAP protein showing GAP activity against both Cdc42 and Rac1 but not RhoA. CdGAP is phosphorylated downstream of the MEK-ERK (extracellular signal-regulated kinase) pathway in response to serum and is required for normal cell spreading and polarized lamellipodia formation. In this study, we found that CdGAP protein and mRNA levels are highly increased in mammary tumor explants expressing an activated Neu/ErbB-2 (Neu-NT) receptor. In response to transforming growth factor-β (TGFβ) stimulation, Neu-NT-expressing mammary tumor explants demonstrate a clear induction in cell motility and invasion. We show that downregulation of CdGAP expression by small interfering RNA abrogates the ability of TGFβ to induce cell motility and invasion of Neu-NT-expressing mammary tumor explants. However, it has no effect on TGFβ-mediated cell adhesion on type 1 collagen and fibronectin. Interestingly, protein expression of E-Cadherin is highly increased in Neu-NT-expressing mammary tumor explants depleted of CdGAP. In addition, complete loss of E-Cadherin expression is not observed in CdGAP-depleted cells during TGFβ-mediated epithelial to mesenchymal transition. Downregulation of the CdGAP expression also decreases cell proliferation of Neu-NT-expressing mammary tumor explants independently of TGFβ. Rescue analysis using re-expression of various CdGAP deletion-mutant proteins revealed that the proline-rich domain (PRD) but not the GAP domain of CdGAP is essential to mediate TGFβ-induced cell motility and invasion. Finally, we found that TGFβ induces the expression and phosphorylation of CdGAP in mammary epithelial NMuMG cells. Taken together, these studies identify CdGAP as a novel molecular target in TGFβ signaling and implicate CdGAP as an essential component in the synergistic interaction between TGFβ and Neu/ErbB-2 signaling pathways in breast cancer cells.     

Clinical usefulness of serum and plasma vascular endothelial growth factor in cancer patients: which is the optimal specimen?

Vascular endothelial growth factor (VEGF) is secreted by various human cancer cells and plays a key role in cancer angiogenesis and metastasis. Recently, evidence of VEGF storage in blood cells including platelets has been reported. The serum VEGF levels were reported to increase during clotting as a result of its release from platelets, and plasma sample instead of serum was recommended for measuring the circulating VEGF more accurately. However, platelets have been implicated in tumor metastasis since circulating tumor cells forming aggregates with platelets were observed. The purpose of this study was to clarify which is an optimal specimen to measure VEGF in cancer patients, serum or plasma. We measured serum and plasma VEGF levels and platelet counts in 173 cancer patients and 42 healthy people, and found that serum VEGF levels were significantly higher than matched plasma VEGF and the VEGF difference (serum VEGF - plasma VEGF) correlated with platelet counts (r=0.624, p<0.05) in both cancer patients and healthy controls. We selected cancer patients with normal platelet counts (130-400x103/microl, Plt-normal cancer group). Interestingly, serum VEGF levels were higher in Plt-normal cancer group than in healthy controls. The theoretical platelet-derived VEGF in serum, calculated based on actual blood platelet counts (pg per 106 platelets), was also significantly higher in Plt-normal cancer group than in normal controls. It is, therefore, suggested that, although the serum VEGF levels are affected by blood platelets, platelet-derived VEGF also reflect biology of cancer cells, and that serum would be the more useful specimen for measurement of circulating VEGF in cancer patients for prognosis.     

Expression of basic fibroblast growth factor, transforming growth factor β1 and their 3 receptors in osteosarcoma and its relationship to angiogenesis

Objective: To investigate the expression of angiogenic factors, basic fibroblast, growth factor (bFGF) and transforming growth factor (TGF)-β₁ in osteosarcoma, its association with neovascularization and prognosis. Methods: The expression of bFGF, β₁ and their receptors, as well as intratumoral microvessel count (MVD) were studied in 80 osteosarcomas by immunohistochemical staining and morphometry. The relationship between the angiogenic factors expression and prognosis was evaluated by a multivariate analysis using Cox proportion hazard model. Results: Among 80 cases of osteosarcoma, 46 cases were positive for TGF/TGF-β (57.5%), and 31 cases for β₁, (RI)(38.8%) respectively. The MVD and bFGF, TGFβ_{1 1} were important indicators to predict the prognosis of patients with osteosarcoma by the Cox proportion hazard model analysis. Conclusion: The angiogenic factors bFGF and TGF-β₁ are involved in the angiogenesis of osteosarcoma, and the angiogenesis influences the prognosis. Also they may be useful in the evaluation of the prognosis of patients with osteosarcoma. This work was supported by a grant from the Youth Scientific Research Foundation of the 9th Five-year Plan of the PLA.     

Mutant epidermal growth factor receptor contributes to head and neck cancer growth and resistance to EGFR targeting

Epidermal growth factor receptor (EGFR) is overexpressed in head and neck squamous-cell carcinoma (HNSCC) and its expression levels correlate with decreased patient survival. Nonetheless, therapies aiming at blocking EGFR has shown limited efficacy in a proportion of patients with HNSCC in clinical trials. Sok et al. in a recent paper (Clin Cancer Res, 2006, 12:5064-5073 ) attempted to ascertain whether it is due to mutation of EGFR. As the most common form of mutation of EGFR seen in several other types of cancer is a truncation mutation, EGFR     

UFT and its metabolite γ-butyrolactone (GBL) inhibit angiogenesis induced by vascular endothelial growth factor in advanced cervical carcinoma

OBJECTIVE: The aim of this study was to evaluate the potential role of UFT and its metabolite gamma-butyrolactone (GBL) for inhibition of angiogenesis induced by vascular endothelial growth factor (VEGF) in advanced cervical carcinoma by the determination of serum GBL and VEGF, and by immunohistochemical staining to assess VEGF protein expression, before and after UFT therapy. METHODS: The subjects were 35 patients with an advanced cervical carcinoma and five healthy volunteers between 2002 and 2003 at Hiroshima University Hospital, under informed consent. The patients received two courses of oral fluoropyrimidine (UFT) therapy at a dose of 600 mg/day for 5 and 2 days off treatment. Serum GBL and VEGF was measured before and after UFT therapy by the gas chromatography mass spectrometry and ELISA-kit in 22 patients and five healthy volunteers, respectively. Immunohistochemical detection of VEGF protein was done in 35 cervical cancers. Results The mean serum GBL level before and after UFT therapy was 21.9 +/- 2.3 and 79.3 +/- 6.2 ng/ml, respectively, and it was significantly increased after UFT administration (P < 0.0001). The mean serum VEGF level before and after UFT therapy was 95.3 +/- 28.1 and 67.5 +/- 11.2 pg/ml, respectively, and it was decreased by UFT administration. In 20 out of 33 (66.6%) patients who were detected with VEGF protein, VEGF protein expression was decreased by UFT therapy. The Delta GBL value (GBL after UFT--GBL before UFT therapy) showed a significant inverse correlation with Delta VEGF value (VEGF after therapy--VEGF before therapy) (r2 = 0.940). CONCLUSIONS: Our findings suggest that UFT and its metabolite GBL inhibit angiogenesis induced by VEGF to have an antitumor effect on cervical cancer.     

Inhibition of hypoxia inducible factor-1α (HIF-1α) decreases vascular endothelial growth factor (VEGF) secretion and tumor growth in malignant gliomas

SummaryIntroduction Hypoxia inducible factor-1α (HIF-1α) regulates vascular endothelial growth factor (VEGF), the presumed principal mediator of angiogenesis in malignant gliomas, under normal physiologic conditions. We examined the effect of HIF-1α on VEGF secretion, tumor growth, and angiogenesis in malignant gliomas.Methods We examined 175 human gliomas for expression of HIF-1α and its downstream-regulated proteins. HIF-1α expression and VEGF secretion in glioma cell lines under normoxia and hypoxia were examined using␣ELISA and Western blot. Malignant glioma cell lines were transfected with dominant-negative HIF-1α (DN-HIF-1α) expression vector or siRNA constructs against the HIF-1α gene. Growth studies were conducted on cells with the highest VEGF/HIF-1α inhibition isolated from stable transfected cell lines. MIB-1-labeling index and microvascular density (MVD) measurements were performed on the in vivo tumors.Results HIF-1 expression correlates with malignant glioma phenotype and was not confined to perinecrotic, pseudopalisading cells. VEGF and HIF-1 expression was high in glioma cell lines even under normoxia, and increased after exposure to hypoxia or growth factor stimulation. Cells transfected with DN-HIF-1α or HIF-1α siRNA demonstrated decreased HIF-1α and VEGF secretion. In vivo but not in vitro growth decreased in response to VEGF and HIF-1 inhibition. HIF-1 siRNA studies showed decreased VEGF secretion and in vitro and in vivo growth of glioma cell lines. MVD was unchanged but MIB-1 proliferation index decreased for both types of HIF-1 inhibition.Conclusions VEGF and HIF-1α are elevated in malignant gliomas. HIF-1α inhibition results in VEGF secretion inhibition. HIF-1α expression affects glioma tumor growth, suggesting clinical applications for malignant glioma treatment.