VEGF In Tissues Research Articles

Protein expression of VEGF, IGF-1 and FGF in retroocular connective tissues and clinical correlation in Graves' ophthalmopathy

To investigate the immunohistochemical expression (IGF-1, EGFr, EGF, c-erbB-2/HER-2/neu, PDGF-A, PDGF-B, FGF and VEGF) in patients with Graves' ophthalmopathy. Twenty-four samples (Graves' ophthalmopathy patients) underwent lateral rectus muscle and surrounding fibrous and adipose tissue biopsy. The control group was obtained by strabismus surgery. Correlation between clinical-ophthalmologic, endocrinological, ultrasonographic findings, and immunohistochemical expression was performed. IGF-1: There were 7 positive cases (29.2%). There was a direct relation with higher CAS (clinical activity score) in all of them and if only CAS equal or higher than 5 was considered, this was 54.5%. FGF: There was expression in 5 cases (20.8%) with a direct relation in all those with higher CAS (>5) (45.4%). VEGF: There were two positive cases (8.3%) for VEGF in endothelial cells, in these cases the patients also presented CAS higher than 5. There was no expressions of all growth factors in the control group. All patients, except one, with positive expression of FGF, IGF-1 and VEGF showed CAS greater than 5, suggesting in this way an important role of these growth factors in the pathogenesis and severity of Graves' ophthalmopathy. However, statistical analysis revealed only significant association between IGF-1 and male sex (P=0.034). Low ultrasound reflectivity and endocrine status may not correlate directly with disease activity or with immunoexpression of growth factors and c-erbB-2/HER-2/neu.

Leukemia inhibitory factor regulates microvessel density by modulating oxygen-dependent VEGF expression in mice

To meet tissue requirements for oxygen, capillaries must be properly distributed without excess or shortage. In this process, tissue oxygen concentration is well known to determine capillary density via the hypoxia-induced cascade, in which HIFs and VEGF play key roles. However, some additional mechanisms modulating this cascade are suggested to be involved in precise capillary network formation. Here, we showed that leukemia inhibitory factor (LIF) was predominantly expressed in developing endothelium, while its receptor was expressed in surrounding cells such as retinal astrocytes. The retinas of Lif(-/-) mice displayed increased microvessel density accompanied by sustained tip cell activity, due to increased VEGF expression by astrocytes in the vascularized area. Lif(-/-) mice resisted hyperoxygen insult in the oxygen-induced retinopathy model, whereas they paradoxically had increased numbers of neovascular tufts. In an in vitro study, LIF inhibited hypoxia-induced VEGF expression and proliferation in cultured astrocytes. Lif(-/-) mice also exhibited similarly increased microvessel density and upregulated VEGF in various tissues outside the retina. Together, these findings suggest that tissues and advancing vasculature communicate to ensure adequate vascularization using LIF as well as oxygen, which suggests a new strategy for antiangiogenic therapy in human diseases such as diabetic retinopathy and cancer.

Expression of endothelial cell-specific adhesion molecules in lungs after cardiac arrest

A method to compensate for donor shortages could be donation after cardiac death. In this study, we considered endothelial cell-specific molecules, claudin-5 and VE-cadherin, as possible biomarkers predicting lung injury against warm ischemia. We investigated how the expression of these molecules could change after cardiac arrest in a mouse lung, comparing other molecules presumably relating with ischemia. At given intervals after cardiac arrest, the lungs were harvested. Quantitative analysis of mRNA expression of claudin-5, VE-cadherin, IL-1beta, IL-10, HIF-alpha, Egr-1, VEGF, Ang-1 and Ang-2 genes in lung tissues with several periods of warm ischemia was performed. Regarding endothelial cell-specific molecules, there were significant differences in both claudin-5 and VE-cadherin mRNA expression between 0 h and 4 h after cardiac arrest. IL-1beta mRNA expression 1 h, 2 h and 4 h after cardiac arrest increased significantly, compared with that at 0 h. There were no significant differences with the other genes. We found that it took more time for claudin-5 and VE-cadherin mRNA expression to change significantly than IL-1beta mRNA expression; therefore, endothelial cell-specific molecules, claudin-5 and VE-cadherin, might be no better candidates for clinical use than IL-1beta.

Expression of VEGF and VEGFR2 in tumors during neoadjuvant therapy of patients with breast cancer

We revealed a significant positive correlation between the concentrations of tissue VEGF and its receptor VEGFR2 in the primary tumor. The degree and direction of changes in the content of VEGF and VEGFR2 in cytosols of tumors from patients with locally spread breast cancer during the preoperation therapy depended on initial levels of these parameters. We found that the expression of the studied factors depended on the degree of therapeutic pathomorphosis in the tumor tissue.

Elevated expressions of survivin and VEGF protein are strong independent predictors of survival in advanced nasopharyngeal carcinoma

BackgroundNasopharyngeal carcinoma (NPC) is one of the most common malignancies in southern China. The China 1992 TNM staging system has been widely used for prognosis prediction of NPC patients in China. Although NPC patients can be classified according to their clinical stage in this system, their prognosis may vary significantly.Method280 cases of NPC with clinical follow-up data were collected and expressions of survivin and VEGF in tumor tissues were investigated by immunohistochemistry (IHC). Apoptosis index (AI) in 100 cases of NPC was detected by the TUNEL method.ResultsExpression of survivin and VEGF were significantly associated with TNM stage, T-stage and metastasis of NPC. The patients with survivin and VEGF over-expression presented lower 5-year survival rate, as compared to those of low-expression (42.32% vs. 70.54%, 40.1% vs. 67.8%, respectively, P < 0.05), especially in advanced stage patients (36.51% vs. 73.41%, 35.03% vs. 65.22%, respectively, P < 0.05). The 5-year survival rate in NPC patients with survivin and VEGF dual over-expression was significantly lower than that of patients with dual low-expression (18.22% vs. 73.54%, respectively; P = 0.0003). Multivariate analysis indicated that both survivin and VEGF over-expression in NPC tumor tissues were strong independent factors of poor prognosis in NPC patients. The mean AI in the 39 survivin low-expression cases was 144.7 ± 39.9, which was significantly higher than that in 61 survivin over-expression cases (111.6 ± 39.8) (T test, P < 0.05).ConclusionSurvivin and VEGF over-expression are independent prognostic factors for the patients with NPC. These results also suggest that tumor survivin and VEGF expressions are valuable prognostic markers for prognosis prediction in NPC patients.

A Role for PPARβ/δ in Ocular Angiogenesis

The uses of highly selective PPARβ/δ ligands and PPARβ/δ knockout mice have shown a direct ability of PPARβ/δ to regulate angiogenesis in vitro and in vivo in animal models. PPARβ/δ ligands induce the proangiogenic growth factor VEGF in many cells and tissues, though its actions in the eye are not known. However, virtually, all tissue components of the eye express PPARβ/δ. Both angiogenesis and in particular VEGF are not only critical for the development of the retina, but they are also a central component in many common pathologies of the eye, including diabetic retinopathy and age-related macular degeneration, the most common causes of blindness in the Western world. This review, therefore, will discuss the recent evidence of PPARβ/δ-mediated angiogenesis and VEGF release in the context of ocular disorders.

Amentoflavone Inhibits Experimental Tumor Metastasis Through a Regulatory Mechanism Involving MMP-2, MMP-9, Prolyl Hydroxylase, Lysyl Oxidase, VEGF, ERK-1, ERK-2, STAT-1, nm23 and Cytokines in Lung Tissues of C57BL/6 Mice

Amentoflavone has been shown to inhibit tumor metastasis in vivo, but its mechanism of action remains unclear. Here, C57BL/6 mice were injected once with B16F-10 melanoma cells via tail vein followed by amentoflavone treatment (50mg/kg BW) for 10 consecutive days. Twenty-one days after tumor injection, animals were euthanized, and tumor metastasis was found to confine in the lungs. As compared with the tumor controls, amentoflavone treatment significantly lowered the number of lung nodules (p<0.001). Amentoflavone treatment markedly decreased the mRNA expression of MMP-2, MMP-9, prolyl hydroxylase, lysyl oxidase, VEGF, ERK-1, ERK-2, TNF-α, IL-1β, IL-6, and GM-CSF in lung tissues. However, amentoflavone treatment increased the mRNA expression of STAT-1 and nm23 in lung tissues. Also in vitro studies indicate that amentoflavone treatment inhibits tumor cell invasion and migration. These results show that amentoflavone treatment reduces experimental tumor metastasis and suggest that such an action is associated with attenuation of tumor invasion, proliferation and angiogenesis.

Expressions and significance of Survivin and VEGF in hepatocellular carcinoma

目的 分析生存素(Survivin)和血管内皮生长因子(VEGF)在原发性肝细胞癌(HCC)中的表达,探讨两者之间的关系.方法 分别应用原位杂交法和免疫组织化学S-P法检测HCC 50例、肝硬化30例、正常肝组织10例中Survivin mRNA、VEGF mRNA和Survivin蛋白、VEGF蛋白的表达情况.结果 Survivin和VEGF在HCC、肝硬化和正常肝组织中的表达差异有统计学意义,Survivin在HCC中的表达高于肝硬化,而VEGF在肝硬化组织中的表达高于HCC中的表达.结论 Survivin和VEGF在HCC中的表达关系密切,呈正相关;Survivin、VEGF的异常表达与HCC的发展密切相关,在HCC的发生、发展过程中起重要的作用。

Prognostic Factors and Recurrence of Small Hepatocellular Carcinoma after Hepatic Resection or Microwave Ablation: A Retrospective Study

PurposeThis study aims to analyze the long-term therapeutic results of small HCC less than 5 cm in diameter after microwave ablation (MA) or hepatic resection (HR) and choose factors that could predict metastasis and recurrence of small HCC. Materials and MethodsThe metastasis and recurrence of 194 patients with one HCC less than 5 cm in diameter who underwent curative HR or MA between January 1995 and December 2004 were reviewed retrospectively; immunohistochemistry was used to analyze the expressions of VEGF, bFGF, and c-Met in HCC tissues. Posttreatment prognostic factors were evaluated by multivariate analysis using Cox’s proportional hazards model. The variables included the expressions of these three proteins in HCC tissues, the clinical and pathologic characteristics of the patients. ResultsThe retrospective study showed that 1-, 3-, and 5-year disease-free survival rates of patients with single HCC of diameter <5 cm were 71.3, 57.0 and 32.5%, respectively. Furthermore, 1-, 3-, and 5-year disease-free survival rates of the patients in MA group and resection group were 72.8, 54.0 and 33.0%; 68.5, 60.0, and 25.6%, respectively. There was no significant difference in disease-free survival rates between these two groups. The result of multivariate analysis showed that differentiation degree of HCC and the expressions of VEGF and c-Met in HCC tissues could be as the independent prognostic factors affecting metastasis and recurrence in patients with small HCC, whereas the methods of therapy had no impact on prognosis. ConclusionsThe metastasis and recurrence rate after MA is similar to that after HR, and the methods of therapy do not affect the prognosis of small HCC. The metastasis and recurrence of patients with small HCC will differ depending on tumor differentiation, expressions of VEGF and c-Met in HCC tissues.

Fibrin‐embedded administration of VEGF plasmid enhances skin flap survival

The aim of the present study was to experimentally evaluate whether topical fibrin-mediated administration of a vascular endothelial growth factor (VEGF)-A plasmid to the wound bed can protect skin flaps from necrosis. A plasmid expression vector containing the VEGF-A cDNA was constructed. The plasmid was then administered to the wound bed of rat abdominal skin flaps in a fibrin sealant. The percentage of viable, ischemic and necrotic tissue was assessed postoperatively as a baseline and after 3 and 7 days using digital surface area morphometry. Laser Doppler imaging of the flaps and VEGF-A Western blot analysis of flap tissue were performed to assess angiogenesis and VEGF-A tissue levels. Flaps treated with VEGF plasmids in the presence of uptake enhancing Lipofectamine transfection reagent increased flap survival 7 days postoperatively significantly associated with markedly elevated tissue perfusion and enhanced tissue VEGF-A protein expression. Our results indicate that topical fibrin-mediated administration of a VEGF-A plasmid may serve as an alternative to previous strategies in treating ischemic skin flaps. The suggested therapeutic approach is easily applicable and inexpensive in preparation. Thus, this protocol may also enhance wound healing in posttrauma skin lacerations or in skin grafts.

Effects of Danshen injection on the malignant obstructive jaundice in the SD rat model

To observe the effects of Danshen on the growth of hepatocellular carcinoma in the SD rats, a model of malignant obstructive jaundice was established by inoculation of transplanted tumor into the hepatic portal with the walker-256 hepatocarcinoma line, which resulted in the obstruction by the infiltration and metastasis of hepatocellular carcinoma. SD rats were divided into 4 groups: the rats were treated by 0.9 % NS (n=24, control group), inosine+vitamin C (n=40, InV group), Danshen (n=40, DS group) and 5-FU (n=40, 5-FU group), respectively. The liver function, morphological changes and the expressions of PCNA, VEGF and ICAM-1 in carcinoma foci, peri-carcinoma tissues, adjacent lobe (left-internal lobe) and lung tissues were observed after the treatment with the 4 agents. Our results showed that the protective effect of Danshen on liver function was significantly better than that of NS and 5-FU (P<0.01). No significant difference in protective effect was observed between DS group and InV group (P>0.05). Danshen also provided protective effect on the morphological damage of liver caused by obstructive jaundice. The rates of carcinoma-inhibition and metastasis inhibition were significantly higher than those of NS and inosine+vitamin C (P<0.01). No significant difference in this regard existed between DS group and 5-FU group (P>0.05). The expressions of PCNA,VEGF and ICAM-1 PCNA, VEGF and ICAM-1 in carcinoma foci, peri-carcinoma tissues, adjacent lobe (left-internal lobe) and lung tissues were lower than those in control group and InV group, with the differences being significant (P<0.01). No significant differences were found between DS group and 5-FU group in the expression levels of PCNA and VEGF (P>0.05) but ICAM-1 (P<0.05). It is concluded that Danshen injection not only has protective effects on liver injury caused by obstructive jaundice, but can inhibit the proliferation and growth of hepatocarcinoma, interfere with the vascularization of tumors, prevent recurrence and metastasis of hepatocarcinoma.

Increases in circulating VEGF levels during COX-2 inhibitor treatment in breast cancer patients

Cyclooxygenase-2 (COX-2) inhibitors are being tried in clinics for cancer treatment. One of the mechanism by which COX-2 inhibitors suppress cancer progression is suggested to be inhibition of angiogenesis. To investigate how COX-2 inhibitors affect regulation of angiogenic factors, we studied alterations in VEGF levels in sera and plasma during COX-2 inhibitor treatment in breast cancer patients. Serum and plasma VEGF levels were monitored in 48 patients treated with the COX-2 inhibitor celecoxib together with 5-FU, epirubicine, cyclophosphamide (FEC). Serum VEGF levels showed decreases on day 3 of the first cycle ( P < 0.0001), followed by increases after 3 weeks ( P < 0.0001). Plasma VEGF levels did not show decreases on day 3 but showed increases after 3 weeks ( P < 0.05). The increases of VEGF levels in sera and plasma continued until the next cycle of the treatment. In patients treated with FEC alone (without celecoxib) did not show increases in serum VEGF levels during the treatment. Our data showed that treatment with COX-2 inhibitors decreased serum VEGF levels at an early time and increased VEGF levels in serum and plasma at a late time in breast cancer patients. Further studies are necessary to elucidate how COX-2 inhibitors regulate production of VEGF in different cells and different tissues in cancer patients.

Association of VEGF expression with poor prognosis among patients with low-grade neuroendocrine carcinoma

4091 Background: Low-grade neuroendocrine carcinomas (LGNET) can arise from neuroendocrine cells throughout the body and have a wide range of aggressiveness. Reliable predictive and prognostic markers of outcome are lacking. Angiogenesis is critical for metastasis and tumor growth beyond a small tumor size and VEGF is a powerful mediator of tumor angiogenesis. Methods: LGNET tissue from 50 patients (24 with local-regional disease, 26 with metastasis) who underwent tumor resection at the University of Texas M.D. Anderson Cancer Center was evaluated for expression VEGF by immunohistochemistry. Chi-square and Fisher’s exact test were used to test the association between study parameters. Kaplan-Meier analysis was used to assess the affect of study parameters on progression free survival (PFS). Results: Strong, weak, and negative VEGF expression was observed in 32%, 54%, and 14% of cases respectively. Larger tumor size was observed among patients with strong VEGF expression. Mean tumor sizes in patients with strong, weak and negative VEGF were 4.8, 2.8, and 2.9 cm. Compared to the group with negative VEGF expression, VEGF (weak/strong) expression was associated with metastasis (14% v 58%; P = .045). The median PFS durations of patients with strong and weak VEGF expression were 29 months and 81 months respectively. With a median follow-up duration of 50 months, the median PFS duration for the group with negative VEGF expression has not been reached. Compared by log rank test VEGF expression was associated with poor PFS (P = .022). Conclusions: This study suggests that tissue VEGF expression is associated with aggressive tumor growth and metastasis among patients with LGNET. VEGF expression may serve as a useful prognostic marker following tumor resection. No significant financial relationships to disclose.

Tissue Endostatin Correlates Inversely with Capillary Network in Rat Heart and Skeletal Muscles

The role of angiostatic factors, including endostatin, in regulating physiological angiogenesis is poorly understood. We used normal adult rats under physiological resting conditions to examine the relationship between tissue endostatin, VEGF, and capillary density (CD) in the heart (high metabolic activity) versus the skeletal muscle (relatively low metabolic activity). The heart (left ventricle, LV) and skeletal muscle (anterior tibialis, AT) were dissected from 12-week-old male Sprague-Dawley rats. Transverse cryosections of LV and AT were stained with FITC-conjugated GS-I-lectin. CD was determined by analysis of randomly acquired digital images of the cryosections using Optimas software. Tissue protein levels of endostatin and VEGF were determined by ELISA assays. Tissue endostatin levels were lower in the LV and higher in the AT (135 +/- 39 vs. 663 +/- 114 pg/mg) and VEGF levels were higher in the LV and lower in the AT (41 +/- 3 vs. 27 +/- 4 pg/mg), respectively (n = 7, P < 0.01). CD in LV and AT were 3632 +/- 428 and 437 +/- 44/mm2, respectively (P < 0.01). We demonstrated that an 8.3-fold greater capillary density is related to a 4.9-fold lower level of tissue endostatin and a 1.5-fold higher level of tissue VEGF in the heart (LV) versus the skeletal muscle (AT) of normal rats under physiological resting conditions. Also, exercise training increased capillary density, decreased tissue endostatin and increased tissue VEGF in the skeletal muscle (AT). These findings suggest that tissue endostatin content correlates inversely with capillary network in the muscle tissues with different metabolic activity, and that tissue endostatin may play a very important role in the metabolic control of angiogenesis under physiological conditions.

KERATINOCYTE GROWTH FACTOR RECEPTOR CORRELATES WITH VENOUS INVASION VIA VASCULAR ENDOTHELIAL GROWTH FACTOR IN PANCREATIC CANCER

Purpose: Keratinocyte growth factor (KGF)/fibroblast growth factor-7 (FGF-7), an angiogenic and mitogenic polypeptide that is mainly synthesized by mesenchymal cells, has been implicated in cancer growth and tissue development and repair. Its actions are dependent on binding to a specific cell-surface KGF receptor (KGFR). In the present study, we investigated the expression and role of KGF and KGFR in human pancreatic cancer tissues. Methods: Immunohistochemically, the expression of KGF, KGFR and VEGF in human pancreatic cancer tissues were examined. Moreover, we evaluated co-expression of KGF and KGFR, and correlation with clinicopathological factors. Result: KGFR was localized in cytoplasm and/or membrane of cancer cells in 22 of the 53 (41.5%) patients, and KGF immunoreactivity was detected in cytoplasm of cancer cells in 18 of the 53 (34.0%) patients. Clinicopathologically, KGF and KGFR expression was highly correlated with venous invasion respectively. Moreover, co-expression of KGF and KGFR correlated significantly with venous invasion, VEGF expression and poor prognosis. Conclusion: These findings suggest that co-expression of KGF and KGFR might play an important role in enhancing venous invasion in pancreatic cancer cases by modulating the capacity of angiogenesis.

Tomato juice prevents senescence-accelerated mouse P1 strain from developing emphysema induced by chronic exposure to tobacco smoke

The senescence-accelerated mouse (SAM) is a naturally occurring animal model for accelerated aging after normal development and maturation. SAMP1 strain was reported to show age-related structural and functional changes in lung and to be a murine model of senile lung. We postulated that aging of lung is an important intrinsic process for development of emphysema and even in a short period of tobacco smoke exposure may be able to generate emphysema. At age 12 wk, SAMP1 inhaled air or 1.5% tobacco smoke (total particulate matter 23.9 mg/m3) through the nose for 30 min/day, 5 days/wk, and for 8 wk. The mean linear intercepts (MLI) and destructive index (DI) of lung were significantly increased [air vs. smoke (means+/-SE); MLI, 68.76+/-0.69 vs. 75.34+/-1.70 microm, P<0.05 and DI, 8.61+/-0.38 vs. 16.18+/-1.54%, P<0.05], whereas no significant changes were observed in SAMR1, control mice that show normal aging. In contrast, smoke-induced emphysema was completely prevented by concomitant ingestion of lycopene given as tomato juice [MLI: smoke with/without lycopene (mean+/-SE), 62.87+/-0.8 vs. 66.90+/-1.33 microm, P<0.05]. Smoke exposure increased apoptosis and active caspase-3 of airway and alveolar septal cells and reduced VEGF in lung tissues, but tomato juice ingestion significantly reduced apoptosis and increased tissue VEGF level. We conclude that SAMP1 is a useful model for tobacco smoke-induced emphysema and a valuable tool to explore both pathophysiological mechanisms and the effect of therapeutic intervention on smoke-induced emphysema.

Modeling prognosis for patients with malignant astrocytic gliomas: Quantifying the expression of multiple genetic markers and clinical variables

The disparate lengths of survival among patients with malignant astrocytic gliomas (anaplastic astrocytomas [AAs] and glioblastoma multiforme [GBM]) cannot be adequately accounted for by clinical variables (patient age, histology, and recurrent status). Using real-time quantitative reverse transcription-polymerase chain reaction, we quantified the expression of four genes that were putative prognostic markers (CDK4, IGFBP2, MMP2, and RPS9) in a set of 43 AAs, 41 GBMs, and seven adjacent normal brain tissues. We previously explicated the expression and prognostic value of PAX6, PTEN, VEGF, and EGFR in these glioma tissues and established a comprehensive prognostic model (Zhou et al., 2003). This study attempts to improve that model by including four additional genetic markers, which exhibited a differential expression (P < 0.001) among tumor grades and between tumor and normal tissues. By including eight log-scaled gene expression variables, three clinical variables, and interaction terms among the eight genes, we established a prognostic model that accounted for two thirds of the variation (R2) in survival for this set of patients. To improve the R2 of the model without compromising its clinical utility, our data demonstrated that incorporating genes from different pathways markedly strengthens the model. Spearman rank correlation analysis of gene expression demonstrated a statistically significant positive correlation (P < 0.01) between the expression of IGFBP2-MMP2 and IGFBP2-VEGF in GBMs, but not in AAs. This finding suggests that the expression of IGFBP2 is associated with pathways activated specifically in GBMs that result in enhancing invasiveness and angiogenesis.

Expression and significance of tumor infiltrating dendritic cells in renal cell carcinoma

Objective: To study the expression of dendritic cells in human renal cell carcinoma and explore the cause, so to reveal the mechanism of escaping immune surveillance in RCC. Methods: The expressions of CD83+DCs, CD1a+DCs,VEGF and TGF-β1 in tumoral, peritumoral and normal kidney tissues of RCC in 30 cases were detected by immunohistochemistry using streptavidin/peroxidese(SP) Results: CD83+DCs were mainly located in the peritumoral areas; whereas CD1a+DCs were mainly retained within the cancer nests. The number of CD83+DCs was inversely correlated with the clinical stage (P 0.05). The expressions of CD83+DCs and CD1a+DCs have significant difference between the tumoral, peritumoral and normal kidney tissues(P 0.05). Conclusion: DC has the tendency to gathering in tumor, but because of the immunosuppressive cytokins, for example VEGF and TGF-β1, inhibits the maturation of DC, there are less mature TIDCs(CD83+TIDCs) in the tumoral tissues, they are mainly located in the peritumoral areas. This may contribute to the mechanism of escaping immune surveillance in RCC.

Vascular Endothelial Growth Factor (Vegf) and Other Common Tissue Prognostic Indicators in Breast Cancer: A Case-Control Study

VEGF is a specific mitogen and survival factor for endothelial cells and a key promoter of angiogenesis in physiological and pathological conditions. Nevertheless, VEGF tissue evaluation in cancer patients as a prognostic factor compared to the conventional histological and biological parameters is still controversial. In this case-control study, tissue VEGF was retrospectively determined by immunohistochemistry and related to T, N, ER, PgR, c-erbB-2, p53, MIB-1 and cyclin D1 in 129 breast cancer patients. Seventy-four of these patients had developed distant metastases postoperatively. The remaining 55 patients had remained disease-free >10 years after surgery. In 17 (13%) of the 129 patients (six with distant metastases and eleven disease-free) tissue and plasma VEGF were concomitantly evaluated. In univariate analysis no significant differences in VEGF and tumor size were found between metastatic and disease-free patients, whereas there were significant differences in N, ER, PgR, c-erbB-2, p53, MIB-1 and cyclin D1 (p ranging from 0.001 to 0.0001). In multivariate analysis VEGF showed less significance than N, ER, c-erbB-2, MIB-1 and cyclin D1 (p = 0.012, p = 0.007, p = 0.005, p = 0.005, p = 0.002 and p = 0.001, respectively). VEGF was a significant unfavorable prognostic indicator only in the N+ subset (p = 0.015), while ER (p = 0.05 and p = 0.021) and MIB-1 (p = 0.031 and p = 0.022) were significant in both the N+ and N- subgroups. In multivariate analysis in the 74 metastatic cases VEGF did not show any significance in relation to disease-free interval and overall survival from the time of mastectomy and from the time of relapse, whereas N and PgR did (p ranging from 0.018 to 0.001). In conclusion, tissue VEGF does not seem a suitable candidate to replace conventional histological and other common biological prognostic factors in breast cancer.

Expression and role of VEGF and neuropilins in pancreatic cancer

Background. Neuropilins (NRPs) are characterized as co-receptors for VEGF. In this study, we determined the expression of NRPs, VEGF, and VEGFRs in pancreatic cancer cells and tissues as well as VEGF-induced cell proliferation. Methods. Human pancreatic cancer cell lines (Panc-1 and MIA PaCa-2), normal human pancreatic ductal epithelium cells (HPDE), and human umbilical vein endothelial cells (HUVECs) were cultured. Human pancreatic adenocarcinoma tissues were also studied. The expression levels of NRPs, VEGFRs, and VEGF were determined by real-time PCR and immunostaining. Cell proliferation was performed by [ 3H]thymidine incorporation. Results. NRP-1 and NRP-2 were both expressed in Panc-1, HPDE, and HUVECs, but minimally expressed in MIA PaCa-2 cells. Panc-1 expressed 30 times higher NRP-1 mRNA than NRP-2 mRNA. NRP-1 mRNA in Panc-1 cells was 5.3 times higher than that in HPDE cells, but similar to that in HUVECs. NRP-2 mRNA in Panc-1 was similar to that in HPDE cells but lower than that in HUVECs. There was no expression of all three VEGFRs in all cell types except for HUVECs. However, the VEGF mRNA was detected in all cell types except for HUVECs. Immunoreactivity of NRP-1 was much higher than that of NRP-2 in Panc-1 and human pancreatic adenocarcinoma tissues, while VEGFRs were not detected. In response of VEGF 165, [ 3H]thymidine incorporation in Panc-1 was significantly increased by 61% ( P < 0.01). Neutralizing antibody against NRP-1 significantly blocked VEGF-induced cell proliferation in Panc-1 cells. Conclusions. Pancreatic cancer cell line Panc-1 and adenocarcinoma tissues expressed high levels of NRP-1 and VEGF, but not VEGFRs, and exogenous VEGF significantly increased Panc-1 cell proliferation. Blocking NRP-1 significantly abolished the effect of VEGF on Panc-1 proliferation. These data suggest that direct interaction between VEGF and NRP-1 may be involved in pancreatic cancer pathogenesis.