Vascular Endothelial Growth Factor Staining Research Articles

Expression of Hypoxia-Inducible Factor (HIF)-1a-Vascular Endothelial Growth Factor (VEGF)-Inhibitory Growth Factor (ING)-4- axis in sarcoidosis patients

BackgroundSarcoidosis is a granulomatous disorder of unknown etiology. The term of immunoangiostasis has been addressed by various studies as potentially involved in the disease pathogenesis. The aim of the study was to investigate the expression of the master regulator of angiogenesis hypoxia inducible factor (HIF)-1a – vascular endothelial growth factor (VEGF)- inhibitor of growth factor 4-(ING4) - axis within sarcoid granuloma.MethodsA total of 37 patients with sarcoidosis stages II-III were recruited in our study. Tissue microarray technology coupled with immunohistochemistry analysis were applied to video-assisted thoracoscopic surgery (VATS) lung biopsy samples collected from 37 sarcoidosis patients and 24 controls underwent surgery for benign lesions of the lung. Computerized image analysis was used to quantify immunohistochemistry results. qRT-PCR was used to assess HIF-1a and ING4 expression in 10 sarcoidosis mediastinal lymph node and 10 control lung samples.ResultsHIF-1a and VEGF-ING4 expression, both in protein and mRNA level, was found to be downregulated and upregulated, respectively, in sarcoidosis samples compared to controls. Immunohistochemistry coupled with computerized image analysis revealed minimal expression of HIF-1a within sarcoid granulomas whereas an abundant staining of ING4 and VEGF in epithelioid cells was also visualized.ConclusionsOur data suggest an impairment of the HIF-1a – VEGF axis, potentialy arising by ING4 overexpression and ultimately resulting in angiostasis and monocyte recruitment within granulomas. The concept of immunoangiostasis as a possible protection mechanism against antigens of infectious origin needs further research to be verified.

Tumor stromal vascular endothelial growth factor A is predictive of poor outcome in inflammatory breast cancer

BackgroundInflammatory breast cancer (IBC) is a highly angiogenic disease; thus, antiangiogenic therapy should result in a clinical response. However, clinical trials have demonstrated only modest responses, and the reasons for these outcomes remain unknown. Therefore, the purpose of this retrospective study was to determine the prognostic value of protein levels of vascular endothelial growth factor (VEGF-A), one of the main targets of antiangiogenic therapy, and its receptors (VEGF-R1 and -R2) in IBC tumor specimens.Patients and MethodsSpecimens from IBC and normal breast tissues were obtained from Algerian patients. Tumor epithelial and stromal staining of VEGF-A, VEGF-R1, and VEGF-R2 was evaluated by immunohistochemical analysis in tumors and normal breast tissues; this expression was correlated with clinicopathological variables and breast cancer-specific survival (BCSS) and disease-free survival (DFS) duration.ResultsFrom a set of 117 IBC samples, we evaluated 103 ductal IBC tissues and 25 normal specimens. Significantly lower epithelial VEGF-A immunostaining was found in IBC tumor cells than in normal breast tissues (P <0.01), cytoplasmic VEGF-R1 and nuclear VEGF-R2 levels were slightly higher, and cytoplasmic VEGF-R2 levels were significantly higher (P = 0.04). Sixty-two percent of IBC tumors had high stromal VEGF-A expression. In univariate analysis, stromal VEGF-A levels predicted BCSS and DFS in IBC patients with estrogen receptor-positive (P <0.01 for both), progesterone receptor-positive (P = 0.04 and P = 0.03), HER2+ (P = 0.04 and P = 0.03), and lymph node involvement (P <0.01 for both). Strikingly, in a multivariate analysis, tumor stromal VEGF-A was identified as an independent predictor of poor BCSS (hazard ratio [HR]: 5.0; 95% CI: 2.0-12.3; P <0.01) and DFS (HR: 4.2; 95% CI: 1.7-10.3; P <0.01).ConclusionsTo our knowledge, this is the first study to demonstrate that tumor stromal VEGF-A expression is a valuable prognostic indicator of BCSS and DFS at diagnosis and can therefore be used to stratify IBC patients into low-risk and high-risk groups for death and relapses. High levels of tumor stromal VEGF-A may be useful for identifying IBC patients who will benefit from anti-angiogenic treatment.

Effect of fibrin on osteogenic differentiation and VEGF expression of bone marrow stromal cells in mineralised scaffolds: a three-dimensional analysis

The aim of the present study was to test the hypothesis that a fibrin matrix enhances the osteogenic differentiation and expression of vascular endothelial growth factor (VEGF) by human bone marrow stromal cells (hBMSCs) seeded into mineralised scaffolds. Porous calcium carbonate scaffolds were droplet seeded with hBMSCs using a matrix containing 3 % fibrinogen and cultured for 3 weeks. Seeded scaffolds without the fibrin matrix served as controls. The scaffolds were evaluated, using undecalcified thick sections, for fluorescence staining for nuclei, osteocalcin (OC) and VEGF. The sections were systematically scanned using optical sectioning and three dimensional distributions of cells and positive staining indicating expression of OC and VEGF were reconstructed from the z-stacks. The fibrin matrix maintained a significantly higher level of cell numbers after 2 d and 1 week and delayed the onset of osteogenic differentiation while sustaining a significantly higher level of OC and VEGF expression after 2 and 3 weeks, starting from the periphery of the scaffolds. There was a decrease in cell density from the periphery to the centre of the scaffolds in both groups The percentage of cells expressing OC and VEGF was significantly different between the centre and the periphery of the scaffolds in the fibrin(+) group but not in the controls. It is concluded that the fibrin matrix used appears to be a useful adjunct for supporting and sustaining osteogenic and angiogenic activity of hBMSCs in tissue engineered constructs. This could help to improve their performance in a clinical setting.

Expression of vascular endothelial growth factor can predict distant metastasis and disease-free survival for clinical stage III rectal cancer following 30-Gy/10-f preoperative radiotherapy

The purpose of this study was to analyze the prognostic value of vascular endothelial growth factor (VEGF) in patients with stage III rectal cancer treated with 30-Gy/10-fraction (f) preoperative radiotherapy and radical surgery. One hundred and sixteen patients with clinical stage III rectal cancer were studied. All patients underwent radical surgery after 30-Gy/10-f preoperative radiotherapy. Immunohistochemical examination of VEGF was performed on the resected tumor specimen. An immunohistochemical score established from the extension and intensity of the markers was used for analysis. The log-rank test and proportional hazards regression analysis were used to calculate the probability that this biomarker was associated with patient outcomes. The expression of VEGF was identified among 74.1% (86 of 116) of patients. We found an increased incidence of distant metastasis (19.8 vs. 3.3%, p = 0.039) and a decreased 3-year disease-free survival rate (96.7% vs. 72.7%, p = 0.003) in patients with positive VEGF staining. In multivariate survival analysis, positive VEGF staining (hazard ratio (HR) = 9.364, 95% confidence interval (CI) 1.298-71.519, p = 0.027) and ypN+ stage (HR = 2.714, 95% CI 1.419-3.331, p = 0.000) were the independent prognostic factors for disease-free survival. Subgroup analyses showed that the expression of VEGF was significantly associated with increased distant metastasis rate and decreased DFS in patients with ypN+ stage. VEGF expression may have potential value for predicting distant metastasis and disease-free survival in stage III rectal cancer after 30-Gy/10-f preoperative radiotherapy. Inhibition of VEGF in combination with radiotherapy may improve the patient outcomes.

The use of molecular markers as a method to predict the response to neoadjuvant therapy for advanced stage rectal adenocarcinoma

The response to combined neoadjuvant therapy for advanced stage rectal adenocarcinoma is predictive of outcome. In addition to both clinical and pathological features, the expression of a variety of molecules may provide another method of identifying tumour responsiveness to pre-operative therapy. The aim of this study was to evaluate several markers in the apoptotic pathway as well as expression of Cox-2 and vascular endothelial growth factor (VEGF) to determine their ability to predict response to neoadjuvant therapy. In total, 152 patients with advanced rectal adenocarcinoma were treated with neoadjuvant therapy followed by resection. Paraffin-embedded sections obtained before and after therapy were assessed by immunohistochemical staining for Cox-2, VEGF, p53, p21, p27, Bax, BCL-2 and apoptosis protease-activating factor 1 (APAF-1). These stains were correlated with tumour regression grade, complete pathological response and T-downstaging of the surgical specimen. Clinical and pathological data were also collected. Data were analysed using the χ2 and Spearman's correlation tests. Pathological complete response was seen in 24.5% of patients. Amongst the apoptosis-associated markers, only APAF-1 expression was found to be significantly associated with tumour regression grade (P<0.001), complete pathological response (P<0.031) and T-downstaging (P<0.004). On multivariate analysis, APAF-1 expression was found to be independently associated with good tumour regression grade. In contrast, overexpression of Cox-2 and VEGF in pretreatment biopsies was related to less tumour regression (P<0.003) and less likelihood of T-downstaging (P<0.03). Immunohistochemical evaluation of initial biopsy specimens of rectal cancer with APAF-1, Cox-2 and VEGF may predict tumour response to neoadjuvant therapy in patients with advanced rectal adenocarcinoma. Those with an expected limited response may be considered for other investigational neoadjuvant protocols.

Effect of two selenium sources on hepatocarcinogenesis and several angiogenic cytokines in diethylnitrosamine-induced hepatocarcinoma rats

This experiment was designed to compare the effect of two selenium sources at the dosage of therapeutic level on hepatocarcinogenesis and angiogenic cytokines in DEN-induced hepatocarcinoma rats to further approach their possible anticancer's mechanism. One hundred and seventy-eight Sprague-Dawley (SD) rats (average weight being 100–120g) were randomly divided into 5 groups (I–V). Animals in group I, group II and group III served as the negative control, sodium selenite control (SS) and positive controls respectively, and received 0.1, 3.0, and 0.1mg/kg selenium from sodium selenite supplemented diets during the whole experimental time. Rats in group IV and group V were fed with selenium from selenium-enriched malt (SEM) and sodium selenite (SS) supplemented diets (3mg/kg respectively). To balance the nutritional content among each group, normal malt which was not treated with selenium was added into the diets of the challenge groups. The nutrition contents, except the selenium of the diet in each group, were similar and in accordance with NRC standards. Rats in groups III–V were treated by aqueous diethylnitrosamine solution (100mg/L) at the dosage of 10mg/kg body weight every day for 16 weeks to induce hepatocarcinoma, and drank sterilized water for an additional two weeks. Rats in group I and group II drank sterilized water throughout the experiment. At 4th, 8th, 12th, 16th week, five rats in each group were then sacrificed by cervical decapitation. At the termination of the study, at 18th week, the surplus rats were sacrificed by cervical decapitation. Feed was withheld from the rats for 12h before sampling. The number of hepatoma nodules in liver and mortality of rats were calculated. The values of the following items, including α-fetoprotein (AFP), gamma-glutamyltranspeptidase (GGT), tumor necrosis factor-α (TNF-α), insulin-like growth factors-II (IGF-II), nitric oxide (NO) and total nitric oxide synthase (T-NOS) in plasma were determined. At the same time, the positive numbers of vascular endothelial growth factor (VEGF) and protein kinase C-α (PKCα) staining cells in tumor tissue were analyzed by immunohistochemistry using the Envision two step methods with a kit. The results indicated that SEM could significantly decrease the mortality of rats and the number of hepatoma nodules, values of GGT and AFP, and the levels of IGF-II, NO and NOS and lessen the positive numbers of VEGF and PKCα staining cells in tumor tissue. Moreover, SEM could increase the levels of TNF-α in the initiated time of hepatocarcinogenesis, whereas, decrease the levels of TNF-α in the progressive time of hepatocarcinogenesis. SS could only significantly inhibit the positive numbers of PKCα staining cells in tumor tissue, decrease the levels of GGT, AFP and TNF-α at minority sampling times, and increase the levels of NO. In conclusion, SEM could reduce the mortality. It might be related to deaden significantly the lesion of liver, delay the cause of hepatocarcinogenesis, and inhibit the progress of angiogenesis to increase the livability of DEN-induced hepatocarcinoma rats. SS at the same therapeutic dosage had less effect on the hepatocarcinogenesis by inhibiting angiogenesis and relative cytokines to some extent.

Low VEGF expression in conceptus material and maternal serum AFP and levels as indicators of defective angiogenesis in first-trimester miscarriages

The aims of this study were to assess the relationship between early miscarriages and vascular endothelial growth factor (VEGF) expression and to determine the serum levels of first-trimester maternal alpha-fetoprotein (AFP) and human chorionic gonadotropin (β-hCG) as markers of angiogenesis and predictors of abortion and intrauterine fetal loss. The present study was a prospective, single-center, randomized controlled clinical trial. Ninety-five women who were 6-10 weeks pregnant between May and June 2010 were included in the study. The subjects were divided into three groups, i.e., incomplete abortion (IA) (n=31), intrauterine death (IU-D) (n=32) and control (elective pregnancy termination) (n=32). Feto-placental materials were compared based on immune staining for VEGF in the pathology laboratory, and maternal serum samples were tested in the hormone laboratory. Serum β-hCG levels in the patient groups were significantly lower than the controls (p=0.001). The serum AFP level was lower than the controls in the IA group while it was higher than the controls in the IU-D (p=0.016). Immunohistochemistry showed that the cytotrophoblast, syncytiotrophoblast and endometrial gland epithelium were weakly stained for VEGF in the patient groups (IA and IU-D) in comparison to the control group (p=0.06, p=0.028, p=0.006). Early pregnancy losses are related to insufficient angiogenesis, and maternal serum AFP and β-hCG can be used as markers of angiogenesis in the first trimester.

Effect of valproic acid treatment on penile structure in prepubertal rats

The aim of this study was to determine the histological effects of valproic acid (VPA) on the penis in prepubertal rats. Twelve male Wistar rats (21-24 days old) were divided equally into 2 experimental groups, and given tap water (control group) or 300mg/kg/day VPA via gavage for 30 days. After the penes had been harvested, the antiangiogenic and antifibrogenic properties of VPA were evaluated immunohistochemically using vascular endothelial growth factor (VEGF), inducible nitric oxide synthase (iNOS), transforming growth factor-beta 1 (TGF-β1) and Masson's trichrome staining. Apoptosis was determined by caspase-3 and caspase-9 immunoreactions. Results were expressed as histochemical score (HSCORE), a semi-quantitative analysis for the intensity of immunohistochemical reactivity. Immunohistochemical HSCORE decreased for VEGF and TGF-β1 staining and increased for iNOS staining in rats treated with VPA compared with the control group. Intensities of caspase-3 and caspase-9 labeling were also significantly increased by administration of VPA. Masson's trichrome staining exhibited a fairly diminished level of collagen in the corpus cavernosum of rats treated with VPA. In the light of these results, the administration of VPA from prepuberty to adulthood led to increased apoptosis and deterioration of the smooth muscle/collagen ratio in rat's corpus cavernosum.

Microvascular density as an independent predictor of clinical outcome in renal cell carcinoma: an automated image analysis study

Tumor microvascular density (MVD) has been shown to correlate with the aggressiveness of several cancers. With the introduction of targeted anti-angiogenic therapy, assessment of MVD has the potential not only as a prognostic but also as a therapeutic marker. The significance of tumor vascularity in clear cell renal cell carcinoma (ccRCC) has been debated, with studies showing contradictory results. Previous studies were limited by manual quantification of MVD within a small area of tumor. Since then, the validity of this method has been questioned. To avoid the inaccuracies of manual quantification, we employed a computerized image analysis, which allowed assessment of large areas of tumor and adjacent normal tissue. The latter was used as an internal reference for normalization. MVD and vascular endothelial growth factor (VEGF) were assessed in 57 cases of ccRCC. Sections were immunostained for CD34 and VEGF. Areas of ccRCC and normal kidney medulla were analyzed within scanned images using software that counted CD34-positive vessels and measured the intensity of VEGF staining. We obtained unadjusted values from tumoral areas and calculated adjusted values as tumor/normal ratios. Unadjusted MVD had no association with clinical outcome. However, similarly to tumor stage, higher adjusted MVD was associated with shorter disease-free survival (log-rank P=0.037, Cox P=0.02). This was significant in univariate and multivariate analyses. MVD did not correlate with tumor stage, pointing to its independent prognostic value. As expected due to the known molecular abnormalities in ccRCC, most tumors showed higher VEGF expression than normal tissue. Higher adjusted VEGF was associated with high tumor grade (P=0.049). The finding of increased MVD as an independent marker of tumor aggressiveness may prove useful in the development of new tests for prognostic and therapeutic guidance. Digital techniques can provide more accurate assessment of immunomarkers and may reveal less obvious associations.

Seasonal Changes in Immunoreactivity of Vascular Endothelial Factor and its Receptors VEGFR1 and VEGFR2 in the Uterus of Wild Ground Squirrels (&lt;i&gt;Citellus dauricus&lt;/i&gt; Brandt)

In this study, we investigated the immunoreactivity of vascular endothelial growth factor (VEGF) and its receptors flt-1 (VEGFR1) and the kinase domain receptor (KDR/Flk-1, VEGFR2) in the uteri of the wild ground squirrels during the estrous period, early pregnancy and nonbreeding period. Cellular localizations of VEGF, VEGFR1 and VEGFR2 were detected by immunohistochemistry, and total proteins were extracted from uterine tissue in the estrous period, early pregnancy and nonbreeding period for Western blotting analysis. In addition, plasma estradiol-17β and progesterone concentrations were measured by radioimmunoassay. Stronger positive staining of VEGF was found in luminal epithelial cells and glandular cells, and its receptors (VEGFR1 and VEGFR2) were observed in stromal cells in the estrous period and early pregnancy compared with the nonbreeding period. The protein levels of VEGF, VEGFR1 and VEGFR2 were significantly higher in the estrous period and early pregnancy as compared with the nonbreeding period. Besides, plasma estradiol-17β and progesterone concentrations were higher in the estrous period and early pregnancy than in the nonbreeding period, suggesting that the immunoreactivities of VEGF, VEGFR1 and VEGFR2 were correlated with changes in plasma estradiol-17β and progesterone concentrations. These results suggested that VEGF and its receptors may be involved in the regulation of seasonal changes in the uterine functions of wild female ground squirrels.

Toll-like receptor 9 and vascular endothelial growth factor levels in human kidneys from lupus nephritis patients

In the lupus mouse and systemic lupus erythematosus (SLE) patients, DNA fragments isolated from plasma may mimic microbial DNA and trigger Toll-like receptor 9 (TLR9) signaling with formation of autoantibodies against DNA fragments and nucleosomes. Vascular endothelial growth factor (VEGF) is a tightly regulated angiogenic cytokine in the kidney. The present study investigated glomerular and tubular expression of both TLR9 and VEGF in biopsies from human subjects with lupus nephritis (LN) and normal controls. Kidney biopsies in LN (n=8) and normal controls (n=10) were evaluated for expression of TLR9 and VEGF. The degree of kidney damage was analyzed according to the International Society of Nephrology / Renal Pathology Society classification. Immunohistochemistry was performed, and slides were incubated with antibodies against VEGF and TLR9 monoclonal antibody, stained with hematoxylin and eosin, mounted and microscopically scored at ×10 and ×20. We observed intense staining of glomeruli and tubules for TLR9 up to 3+ from patients with LN. Samples from LN subjects showed 3+ staining of glomeruli but only up to 2+ in tubules for VEGF. There was less significant staining for TLR9 and none for VEGF in controls. There was no correlation observed between LN class severity and intensity of staining for VEGF or TLR9. This is the first study that investigated combined expression of TLR9 and VEGF, which could be an important tool for understanding the role of TLR9 and VEGF in LN, with insights into the early detection and targeted treatment of this disease.

Expression of RECK in endothelial cells of glioma: comparison with CD34 and VEGF expressions

Angiogenesis is thought to be involved in the progression of glioma grades, and reversion-inducing cysteine-rich protein with Kazal motifs (RECK) has been said to be involved in maturation of vessels. In this study, we aimed to determine whether high micro-vessel density (MVD) expressed by RECK in glioma tissue is correlated with grades of glioma. We also compared RECK expression with that of the formerly known vessel marker, CD34, and vascular endothelial growth factor (VEGF). RECK, CD34, and VEGF immuno-reactivities of 72 glioma tissues were studied. RECK was seen in microvessels of glioma tissues. CD34 showed a similar pattern to RECK, whereas VEGF showed positive staining in cytoplasm of tumor cells and endothelial cells. Average MVD with RECK was 107.6 microvessels (range 7-290). RECK was positively correlated with grades of glioma. RECK and CD34 also showed a strong correlation (P = 0.001). Higher frequency of VEGF staining was also correlated with higher grade of glioma. This is the first study describing expression of RECK in glioma, and its angiogenesis-related nature may provide a potential therapeutic target for glioma treatment in the future.

Inhibitory Effect of Intravitreal Injection of Bevacizumab on Nerve Growth Factor

Purpose: To investigate the inhibitory effect of intravitreal injection of bevacizumab on the expression of nerve growth factor (NGF) in rabbit retina.Methods: The right eyes of 40 New Zealand white rabbits were injected with 1.25 mg (0.05 cc) bevacizumab three times monthly; as controls, the left eyes received sham injections. Apoptosis in retinal cells was evaluated by immunohistochemical staining for caspase-3, and by in situ terminal deoxynucleotidyl transferase-mediated biotin-deoxyuridine triphosphate nick-end labeling (TUNEL) of DNA fragments. NGF and vascular endothelial growth factor (VEGF) proteins in rabbit retinas were measured quantitatively (Enzyme linked immunosorbent assay (ELISA)) and qualitatively (immunohistochemical staining) 1 and 4 months after injection. NGF and VEGF messenger RNAs in rabbit retinas were evaluated by real-time polymerase chain reaction (RT-PCR).Results: As shown by the TUNEL assay and caspase-3 immunostaining, the bevacizumab-injected group had significantly more apoptotic activity than did the control group. Levels of retinal NGF and VEGF proteins in the bevacizumab group were lower than those in the control group (p < 0.05). Immunohistochemical staining of NGF and VEGF was weaker in the bevacizumab group than in the control group. NGF and VEGF mRNA levels in the bevacizumab group were lower than those of the control group (p < 0.05).Conclusions: Findings of the present study suggest that apoptosis in retinal cells after intravitreal bevacizumab injection is increased by down-regulated NGF, caused by VEGF inhibition in rabbits.

Targeting Angiogenesis in Gastroesophageal Cancer: Industry‐Sponsored Trials are not the Answer

Angiogenesis is a dynamic process essential for tumor growth. Maeda et al. first reported its association with poor prognosis in gastric cancer in 1996 and, since then, there have been numerous similar studies examining various factors that mediate angiogenesis such as vascular endothelial growth factor (VEGF) and platelet-derived growth factor [1]. It is therefore surprising that a decade and a half later there is still no clear consensus on the role of antiangiogenic agents in the treatment of gastroesophageal cancer and, more importantly, recognition of the patient subset who are most likely to benefit from these expensive agents. In this issue of the Journal, Liu et al. [2] report that VEGF expression is an independent prognostic factor for tumor recurrence and decreased survival in patients with node-negative gastric cancer. Unlike prior studies, the authors excluded all node-positive patients in the hopes of stratifying node-negative patients into lowand high-risk groups for tumor recurrence. This is an important step in identifying patients most likely to benefit from adjuvant therapy, but the manner in which patients were deemed node-negative is particularly relevant. What was the median number of nodes resected, and how were the nodes examined? It seems unusual that 11 patients with pT4 tumors were node-negative. If we presume all nodes were examined using a single section and conventional [hematoxylin-eosin (H&E)] staining, then this means that only about 1% of the lymph node tissue was examined, and some node-positive patients almost certainly were missed. In a retrospective study last year, we found that almost one-third of esophageal cancer patients who were node-negative by conventional histology proved to have occult tumor deposits in their lymph nodes after serial sectioning and immunohistochemical evaluation [3]. In other words, until standardized and more stringent histopathology is performed on lymph nodes, it is difficult for the reader to be confident that all patients were truly nodenegative. Also, in Liu et al.’s study, it is possible that 24 of 55 (44%) patients who were VEGF-positive and had a worse survival were those who might have been nodepositive with more stringent histopathologic testing. Nevertheless, their study raises an interesting question. Should patients with early gastric cancer and positive VEGF staining be given an antiangiogenic agent to reduce their risk of tumor recurrence? Bevacizumab (Avastin), a recombinant humanized monoclonal antibody directed against VEGF-A, was approved for patients with metastatic colorectal cancer in 2004, in combination with 5-fluorouracil chemotherapy, after a Phase III (randomized controlled multicenter) study showed an increase in survival of 4.7 months with the inclusion of this agent in the treatment regimen [4]. This study paved the way for a myriad of other clinical trials evaluating more than 40 antiangiogenesis agents in various solid tumors. In gastric cancer, a Phase III trial called AVAGAST (Avastin in Gastric Cancer) has just been published in the Journal of Clinical Oncology on the back of three Phase II (nonrandomized) trials evaluating the addition of bevacizumab to chemotherapy in patients with unresectable or metastatic gastroesophageal cancer [5]. Although the smaller Phase II trials reported response rates of 65 to 68% with encouraging progression-free survival [6], the Phase III trial found only a 2 month survival benefit between groups, which was not significant [5]. Subsequently, bevacizumab has not been S. K. Thompson (&) Department of Surgery, Royal Adelaide Hospital, University of Adelaide, Level 5, Eleanor Harrald Building, Adelaide, South Australia 5000, Australia e-mail: sarah.thompson@adelaide.edu.au

Immunohistochemical Study of Vascular Endothelial Growth Factor (VEGF) and Matrilysin (MMP-7) in T1 Adenocarcinoma of the Colon and Rectum to Predict Lymph Node Metastases or Distant Metastases

Predicting the lymph node metastatic or distant metastatic potential of T1 adenocarcinoma of the colon and rectum remains a major challenge. We investigated the role of the expressions of tumor matrilysin (MMP-7), VEGF-C and VEGF-A in predicting these metastatic potentials. Single T1 adenocarcinomas were examined and pathological tumor factors were reviewed. Immunohistochemical staining of VEGF and MMP-7 was performed and the metastatic potential was defined on the basis of the presence of lymph nodes in the specimen or the identification of other distant metastasis during follow-up examinations. There was little correlation between the IHC staining results of VEGF-A, VEGF-C and MMP-7 in the same specimen (kappa<0.1). After a 61-month median follow-up (2-131 months), 17 (11.8%) tumors showed metastatic potential, including 14 lymph node metastases and 3 distant metastases. The tumors showing high levels of MMP-7 expression had higher metastatic potentials than those with low expression (19.1% vs. 8.2%, respectively; p=0.057). Overexpression of MMP-7 generally indicated an inferior overall survival (p=0.09). In analysis of the traditional pathological tumor factors, only lymphovascular invasion showed significance in predicting metastatic potential (p=0.04). Overexpression of MMP-7 generally indicated a higher metastatic potential and inferior overall survival. Lymphovascular invasion was a significant risk factor for lymph node metastasis.

The role of vascular endothelial growth factor in fractional laser resurfacing with the carbon dioxide laser

The aim of this study was to analyze the role of vascular endothelial growth factor (VEGF) in mechanisms of cutaneous remodeling induced by fractional CO(2) laser treatment. The dorsal skin of Kunming mice was exposed to a single-pass fractional CO(2) laser treatment. Biopsies were taken 1h, and 1, 3, 7, 14, 28 and 56days after treatment. Skin samples VEGF expression was evaluated by immunohistochemistry and ELISA, fibroblasts by hematoxylin-eosin staining, and types I and III collagen by ELISA. Staining for VEGF was found in many types of cell including fibroblasts. The amount of VEGF in the skin of laser-treated areas had increased significantly compared to that in the control areas on days 1and 3(P < 0.05, P < 0.01, respectively), then decreased by day 7 after treatment and returned to the baseline level. The number of fibroblasts in the skin of the laser-treated areas had increased significantly compared to that in control areas on days 3, 7, 14, 28 and 56 after irradiation (P < 0.05, P < 0.01, P < 0.01, P < 0.01, P < 0.01, respectively). The amount of type I collagen was significantly higher in the skin of the laser-treated areas compared to that in control areas from day 28 to day 56 (P < 0.05, respectively), and type III collagen was significantly higher from day 3 to day 56 (P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.01, respectively). There was a positive correlation between the level of VEGF and fibroblast proliferation early stage after laser treatment (r = 0.853, P < 0.01), but there was no correlation after the first week (r = -0.124, P > 0.05). The amounts of type I and III collagen showed no significant correlations with the expression of VEGF in the late stages after laser treatment (r = 0.417, P > 0.05 and r = 0.340, P > 0.05, respectively). The results suggest that VEGF might be mainly involved in the early stages of wound healing, including the stages of acute inflammation, fibroblast proliferation and vessel formation induced by fractional CO(2) laser resurfacing.

Prognostic Value of Epithelial Growth Factor Receptor, Vascular Endothelial Growth Factor, E-Cadherin, and p120 Catenin in Resected Non-Small Cell Lung Carcinoma

Introduction Several markers have been investigated to predict the prognosis of lung cancer. In the present study, the prognostic values of epithelial growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), E-cadherin, and p120 catenin expression were investigated by immunohistochemistry in patients with a surgically resected non-small cell lung carcinoma (NSCLC). Patients and method EGFR, VEGF, E-cadherin, and p120 catenin expression were prospectively determined in resected specimens from patients with NSCLC who had undergone surgery between 2003 and 2007. Patients’ and disease-related general characteristics and survival rate were recorded. Results One hundred seventeen patients with a mean age of 61.3 years were included in the study. After a mean follow-up of 27.5 months, the median survival was determined to be 44.0 months and the 5-year survival was 46.2%. The 5-year survival in negative and positive staining groups were as follows; 32% and 66.7% for EGFR ( P=.02), 37.8%, and 50.7% for VEGF ( P=.5), 41% and 66% for E-cadherin ( P=.19), and 46% and 50% for p120 catenin ( P=.27). The differentiation, N status, stage, and EGFR staining were variables significantly affecting survival ( P=.001, .006, .03, and .02, respectively). In multivariate Cox analysis, the EGFR staining level and N status were variables those significantly affecting survival ( P=.021 and P=.010). Conclusions While negative staining of EGFR was related with poor survival, staining of VEGF, E-cadherin, and p120 catenin were not related with survival in patients with resected NSCLC.

Significance of heparanase-1 and vascular endothelial growth factor in adrenocortical carcinoma angiogenesis: potential for therapy

The purpose of this study was to determine the correlation between human adrenocortical carcinoma (ACC) and the proteins involved in tumor angiogenesis, and to evaluate the angiogenic status of ACC. The expression of heparanase-1 (HPA-1), vascular endothelial growth factor (VEGF), and vascular endothelial growth factor receptor-2 (VEGFR-2) as well as microvessel density (MVD) were measured in a series of tissue samples from 44 human sporadic adrenocortical tumors by immunohistochemistry. These specimens were classified as adenomas (n=20) and carcinomas (n=24) according to the histological criteria defined by Weiss. A total of 22 of 24 (91.67%) malignant cases showed positive staining for HPA-1 and 3 of 20 (15%) benign cases showed positive, the difference of HPA-1 expression between ACA and ACC was statistically significant (P<0.001). Similarly, VEGF staining was seen in 70.83% (17/24) of the malignant cases versus 25% (5/20) of the benign, the difference of VEGF expression among two groups was statistically significant (P=0.002). VEGFR-2 expressed highly in the ACC group (79.17%, 19/24) and lowly in the benign group (25%, 5/20), the two groups had extremely significant difference (P<0.001). Malignant cases showed higher MVD compared to benign tumors (84.70±12.44 vs. 21.05±8.07, P<0.001). HPA-1 and VEGF expression were positively correlated with MVD in all specimens (r_s=0.812, P=0.001; r_s=0.834, P<0.001). In conclusion, these results suggest that angiogenesis of human ACC maybe mediated by these proteins and they could represent selective targets for the molecularly targeted treatments of ACC.

Vascular endothelial growth factor expression up-regulated by endometrial ischemia in secretory phase plays an important role in endometriosis

Primary human endometrial cells were exposed to hypoxia preconditioning (HPC), HPC-hypoxia, and hypoxia conditions, and then endometrial tissue treated with ischemia preconditioning (IPC) was transplanted onto the chick embryo chorioallantoic membrane to investigate the role of slight ischemia of endometrium in the pathologic process of endometriosis. IPC up-regulated vascular endothelial growth factor expression and decreased apoptosis of endometrial cells, thus facilitating the endometrial fragments' ectopic implantation.

Phase II randomized trial of bevacizumab versus bevacizumab and thalidomide for relapsed/refractory multiple myeloma: a California Cancer Consortium trial

Vascular endothelial growth factor (VEGF) is upregulated in multiple myeloma (MM), and circulating VEGF levels may correlate with response to therapy (Hideshima, et al 2005, Pittini, et al 2002). Thalidomide has been part of the standard treatment for MM and is thought to inhibit VEGF-associated angiogenesis (Du, et al 2004). Bevacizumab, a monoclonal antibody directed against VEGF-A, inhibits VEGF (Jenab-Wolcott and Giantonio 2009). Accordingly, we set out to test the efficacy and safety of bevacizumab alone and in combination with thalidomide in MM patients.