BACKGROUND: Vascular endothelial growth factor (VEGF)-C and VEGF-D are novel growth factors that regulate lymphatic vessel growth. This study was designed to examine whether the expression of three VEGF family members, VEGF-A, VEGF-C and VEGF-D are associated with the clinicopathologic parameters, especially with lymph node metastasis, in advanced gastric carcinomas. METHODS: Immunohistochemical staining was performed for VEGF-A, VEGF-C, and VEGF-D in the surgically resected specimens from 102 patients with advanced gastric carcinoma. The mRNA expressions of the three VEGF family members were assessed in 16 cases of tumor tissues and their corresponding non-neoplastic tissues. RESULTS: Of the 102 gastric carcinomas, 74 (73%), 82 (80%), and 34 (33%) cases showed cytoplasmic immunoreactivity for VEGF-A, VEGF-C and VEGF-D, respectively. Both VEGF-A and VEGF-C expressions were associated with lymphatic invasion and lymph node metastasis (p<0.05), but the VEGF-D expression was not associated with them (p>0.05). In the tumor tissue, VEGF-C mRNA expression was greater, while VEGF-D mRNA expression was lower than in the nonneoplatic tissue adjacent to the tumor. CONCLUSIONS: VEGF-A and VEGF-C may play important roles for the lymphatic spread of gastric carcinoma. We suggest that neutralizing both VEGF-A and VEGF-C may be reguired to block lymph node metastasis.
Adenocarcinoma* ; Cytoplasm ; Humans ; Intercellular Signaling Peptides and Proteins ; Lymph Nodes ; Lymphatic Vessels ; Neoplasm Metastasis ; RNA, Messenger ; Vascular Endothelial Growth Factor A* ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factor D ; Vascular Endothelial Growth Factors*

PURPOSE: Lymphatic spread of tumor is an important prognostic factor for patients with non-small cell lung carcinoma (NSCLC). Vascular endothelial growth factor-C (VEGF-C) and VEGF-D play important roles in lymphangiogenesis via the VEGF receptor 3 (VEGFR-3). We sought to determine whether VEGF-C, VEGF-D and VEGFR-3 are involved in the clinical outcomes of patients with resected NSCLC. MATERIALS AND METHODS: Using immunohistochemical staining, we investigated the protein expressions of VEGF-C, VEGF-D and VEGFR-3 in the tissue array specimens from patients who underwent resection for NSCLC. The immunoreactivity for p53 was also examined. The clinicopathological implications of these molecules were statistically analyzed. RESULTS: Analysis of a total of 118 specimens showed that VEGF-C, VEGF-D and their co-expression were significantly associated with more advanced regional lymph node metastasis (p=0.019, p=0.044 and p=0.026, respectively, N2 versus N0 and N1). A VEGFR-3 expression had a strong correlation with peritumoral lymphatic invasion (p=0.047). On the multivariate analysis for survival and recurrence, pathologic N2 lymph node metastasis was the only independent prognostic factor, but none of the investigated molecules showed any statistical correlation with recurrence and survival. CONCLUSIONS: The present study revealed that high expressions of VEGF-C and VEGF-D were strongly associated with more advanced regional lymph node metastasis in patients with resected NSCLC.
Carcinoma, Non-Small-Cell Lung ; Humans ; Lung ; Lymph Nodes ; Lymphangiogenesis ; Multivariate Analysis ; Neoplasm Metastasis ; Receptors, Vascular Endothelial Growth Factor ; Recurrence ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factor D ; Vascular Endothelial Growth Factor Receptor-3 ; Vascular Endothelial Growth Factors

OBJECTIVE: Cellular, animal, and human epidemiological studies suggested that benzodiazepines increase the risk of cancer and cancer mortality. Obesity is also clearly linked to carcinogenesis. However, no human studies have examined benzodiazepine-associated carcinogenesis as assessed by changes in cancer biomarkers. METHODS: A total of 19 patients were recruited, and received a 6-week treatment of 0.5 mg lorazepam. The measured cancer biomarkers were angiopoietin-2 (ANG-2), soluble CD40 ligand, epidermal growth factor, endoglin, soluble Fas ligand (sFASL), heparin-binding EGF-like growth factor (HB-EGF), insulin-like growth factor binding protein, interleukin (IL)-6, IL-8, IL-18, plasminogen activator inhibitor (PLGF), placental growth factor, transforming growth factor (TGF)-α, tumor necrosis factor (TNF)-α, urokinase-type plasminogen (uPA), vascular endothelial growth factor (VEGF)-A, VEGF-C, and VEGF-D. RESULTS: Six cancer biomarkers were significantly increased in all patients as a whole. The subgroup analysis revealed a distinct pattern of change. Overweight patients showed a significant increase in 11 cancer biomarkers, including ANG-2, sFASL, HB-EGF, IL-8, PLGF, TGF-α, TNF-α, uPA, VEGF-A, VEGF-C, and VEGF-D. However, normal-weight patients did not show any changes in cancer biomarkers. CONCLUSION: Adiposity may have primed the carcinogenic potential, leading to lorazepam-associated carcinogenesis in overweight patients. Epidemiological studies addressing this issue should consider the potential modulator contributing to benzodiazepine-associated carcinogenesis.
Adiposity ; Angiopoietin-2 ; Animals ; Benzodiazepines ; Biomarkers, Tumor ; Carcinogenesis* ; Carrier Proteins ; CD40 Ligand ; Epidemiologic Studies ; Epidermal Growth Factor ; Fas Ligand Protein ; Heparin-binding EGF-like Growth Factor ; Humans ; Interleukin-18 ; Interleukin-8 ; Interleukins ; Lorazepam ; Mortality ; Obesity ; Overweight* ; Plasminogen ; Plasminogen Activators ; Transforming Growth Factors ; Tumor Necrosis Factor-alpha ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factor D

PURPOSE: CD105 (endoglin) has been shown to be a more useful marker to identify the proliferating endothelium involved in tumor angiogenesis than are the panendothelial markers. The monoclonal antibody D2-40 is a specific lymphatic endothelial marker. METHODS: We investigated CD105, lymphatic vessel marker (D2-40), vascular endothelial growth factor (VEGD)-A and the VEGF-D expressions as possible prognostic markers in the endoscopic biopsy tissue of stomach cancer patients. The pre-operative endoscopic biopsies and surgical biopsies from 73 patients were immunostained for CD105, D2-40, VEGF-A and VEGF-D. Positively stained microvessels were counted in densely vascular foci (hot spots) at a x200 field in each specimen. RESULTS: The microvessel density (MVD) and lymphatic vessel density (LVD), according to the CD105 and D2-40 expressions of the endoscopic biopsies, showed a statistically significant correlation with the surgical biopsies. The MVD via CD105 a showed statistically significant correlation with the histologic differentiation, T-stage, nodal metastasis and stage in the endoscopic biopsies and surgical biopsies, respectively. The lympathic vessel density (LVD) via D2-40 showed a statistically significant correlation with T-stage, nodal metastasis and stage in the endoscopic biopsies. The expressions of VEGF-A and VEGF-D showed a statistically significant correlation with the MVD and LVD. CONCLUSION: The MVD, as determined by the CD105 expression and the LVD as determined by the D2-40 expression may be useful markers for predicting the invasiveness with using a pre-operative endoscopic biopsy of stomach cancer.
Biopsy* ; Endothelium ; Humans ; Lymphatic Vessels ; Microvessels ; Neoplasm Metastasis ; Stomach Neoplasms* ; Stomach* ; Vascular Endothelial Growth Factor A* ; Vascular Endothelial Growth Factor D ; Vascular Endothelial Growth Factors*

<b>OBJECTIVEb>To investigate the relationship between vascular endothelial growth factor C (VEGF-C) expression, VEGFR-3 expression, lymphangiogenesis and angiogenesis in human non-small cell lung cancer (NSCLC).

<b>METHODSb>Seventy-six NSCLC samples were stained for VEGF-C, VEGFR-3 and CD34 with immunohistochemical methods. Assessment of lymphatic vessel density (LVD) and microvessel density (MVD) was performed. The expressions of VEGF-C in 24 fresh NSCLC samples were determined with Western blot assay.

<b>RESULTSb>Of the 76 NSCLC cases, 55 were VEGF-C positive and 40 were VEGFR-3 positive in cancer cells. A significant positive correlation was found between VEGF-C expression and VEGFR-3 expression in cancer cells (P < 0.05). VEGF-C expression was negatively associated with differentiation of tumor cells (P < 0.05). VEGF-C expression and VEGFR-3 expression were positively associated with lymph node metastasis and lymphatic invasion (P < 0.05). LVD was positively related to VEGF-C expression, lymph node metastasis, lymphatic invasion and clinical stage (P < 0.05). There was a significant correlation between LVD and MVD (R = 0.732, P < 0.05). Patients with positive VEGF-C expression had worse outcomes than those with negative VEGF-C expression (P < 0.01).

<b>CONCLUSIONSb>In NSCLC, VEGF-C and VEGFR-3 are related to the lymphangiogenesis, angiogenesis, and occurrence and development of lung cancers. VEGF-C expression could be a useful predictor of poor prognosis in NSCLC.

Carcinoma, Non-Small-Cell Lung ; metabolism ; Endothelial Growth Factors ; biosynthesis ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factor Receptor-3 ; biosynthesis

<b>OBJECTIVEb>To investigate the expression of VEGF-C and VEGF-D and their correlations with lymphangiogenesis and angiogenesis in gallbladder carcinoma.

<b>METHODSb>Fifty cases of gallbladder carcinoma with complete clinical and pathological data were analyzed. The expression of VEGF-C and -D, D2-40, CD31 was assayed by immunohistochemical staining, with 10 samples of normal gallbladder tissues away from cancer and 19 samples of chronic cholecystitis as controls, and their correlation with clinicopathological findings were analyzed retrospectively.

<b>RESULTSb>Thirty-two (64.0%) of the 50 gallbladder cancers were positive for VEGF-C protein expression by immunohistochemistry and the positive rate of VEGF-D protein expression was 62.0% (31/50). The protein expression of VEGF-C and VEGF-D in tumor tissues was significantly higher than that in normal gallbladder tissues away from the tumor (P < 0.05), but no correlation with that in chronic cholecystitis (P < 0.05). The VEGF-C expression correlated with the patient age and lymph node metastasis (both P < 0.05). The VEGF-D expression only correlated with lymph node metastasis (P < 0.05). In the 50 gallbladder cancers, the MLVD was 6.9 + or - 3.6 and the MVD was 36.1 + or - 12.8. The MLVD in both VEGF-C and -D positive groups was significantly higher than that in the negative groups (P = 0.000), and the lymph node metastasis also increased. MVD in both VEGF-C and -D positive groups was higher than that in the negative groups (P < 0.05), and it was also correlated with tumor differentiation (P < 0.05). A significant positive correlation was also found between VEGF-C and VEGF-D expression (r = 0.498, P < 0.01).

<b>CONCLUSIONb>VEGF-C and VEGF-D are involved in the lymphangiogenesis and angiogenesis in gallbladder carcinoma, promote lymph node metastasis of the tumor, and both are important in the regulation of lymphangiogenesis and angiogenesis in this cancer. VEGF-C and VEGF-D are of clinical significance in evaluating lymph node metastatic potency and estimation of prognosis in gallbladder carcinoma.

Adenocarcinoma ; blood supply ; metabolism ; pathology ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Carcinoma, Papillary ; blood supply ; metabolism ; pathology ; Cholecystitis ; metabolism ; pathology ; Female ; Gallbladder Neoplasms ; blood supply ; metabolism ; pathology ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; Retrospective Studies ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVES: To investigate the relationships between lymph node metastasis (LNM) and expression of CD31, D2-40 and vascular endothelial growth factors (VEGF)-A and -C in patients with papillary thyroid cancer (PTC). METHODS: Paraffin-embedded thyroid tissues of 72 patients were evaluated, which included 25 patients with thyroid nodular hyperplasia (TNH), 24 PTC patients without LNM, and 23 PTC patients with LNM. Three pathologists, who were blinded to the patient's clinical information, assessed the immunohistochemical staining results. The amount of expression was scored as high (>25% of cells stained) or low (0-25%). RESULTS: A higher level of VEGF-A expression was observed in the PTC groups regardless of LNM when compared to the group with TNH (91.3%, 79.2%, 4.0%, respectively). VEGF-C expression in the PTC with LNM group was significantly higher than the other two groups (P<0.05). No difference in microvessel density (MVD) scores was observed using CD31 among the three groups. The lymphatic vessel density (LVD) score using D2-40 was significantly higher in patients having PTC with LNM than the other groups (P<0.05). CONCLUSION: VEGF-C and D2-40 were more highly expressed in patients having PTC with LNM than in patients having PTC without LNM or in those having TNH. Analysis of VEGF-C level and LVD using D2-40 may be helpful in the diagnosis of PTC and the evaluation of LNM potential in patients with PTC.
Carcinoma, Papillary ; Factor IX ; Glycosaminoglycans ; Humans ; Hyperplasia ; Lymph Nodes ; Lymphatic Vessels ; Microvessels ; Neoplasm Metastasis ; Thyroid Gland ; Thyroid Neoplasms ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factors

PURPOSE: Vascular endothelial growth factor (VEGF) is one of the most important growth factors for metastatic tumors. To clarify the role of VEGF-A and C in patients with peptic ulcer disease (PUD) or gastric cancer (GC), we evaluated the expression levels of these two molecules. We also analyzed the effect of Helicobacter pylori infection on VEGF-A and C expression levels. MATERIALS AND METHODS: Patients with dyspepsia who needed diagnostic endoscopy were selected and divided into three groups: non-ulcer dyspepsia (NUD), PUD, and GC, according to their endoscopic and histopathological results. Fifty-two patients with NUD, 50 with PUD, and 38 with GC were enrolled in this study. H. pylori infection was diagnosed by the rapid urease test. After RNA extraction and synthesis of cDNA, the expression levels of VEGF-A and C were determined by quantitative reverse transcriptase polymerase chain reaction. RESULTS: The VEGF-C expression level in the PUD and GC groups was significantly higher than that in the NUD group. Moreover, the VEGF-A expression level in the PUD and GC groups was higher than in the NUD group, although the differences were not statistically significant. Significant positive correlations were also observed between the expression levels of these two molecules in the PUD and GC groups. In addition, the expression levels of these two molecules were higher in H. pylori positive patients with PUD or GC than in H. pylori negative patients of the same groups; however, these differences did not reach statistical significance. CONCLUSIONS: Up-regulation of VEGF-C expression during gastric mucosal inflammation may play a role in the development of peptic ulcers or GC.
DNA, Complementary ; Dyspepsia ; Endoscopy ; Helicobacter pylori ; Humans ; Inflammation ; Intercellular Signaling Peptides and Proteins ; Peptic Ulcer* ; Reverse Transcriptase Polymerase Chain Reaction ; RNA ; Stomach Neoplasms* ; Up-Regulation ; Urease ; Vascular Endothelial Growth Factor A* ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factors*

growth factors including VEGFR-2, VEGFR-3. E7080 is an orally active inhibitor of multiple tyrosine kinases including VEGFR-2, 3. Therefore, it was proposed that E7080 may inhibit angiogenesis and lymphangiogenesis. The aim of this study was to determine the effect of E7080 in a nude mouse model of OSCC.METHODS: KB cells were xenografted into the submucosal tissue of the mouth floor of athymic mice. Seven days after the xenograft, the mice were randomized into 2 groups. E7080 were administered orally to the experimental group once per day. The mice were sacrificed 3 weeks after the treatment. The tumors were examined histopathologically. Immunohistochemical assays with anti- VEGF-C, VEGFR-2, VEGFR-3, phosphorylated VEGFR-2/3 (pVEGFR-2/3), and D2-40 antibodies were then performed.RESULTS: The transplantation of human OSCC tumor cells into the mouth floor resulted in the formation of orthotopic tumors. The experimental (E7080 treatment) group showed a slowly increased tumor volume. Moreover, immunohistochemical staining demonstrated higher levels of VEGF-C, VEGFR-2, VEGFR-3, pVEGFR-2/3 and D2-40 expression in the control group than in the experimental group.CONCLUSION: These results suggest that E7080 may provide therapeutic benefits in OSCC.]]>
Animals ; Antibodies ; Carcinoma, Squamous Cell ; Endothelial Growth Factors ; Humans ; Intercellular Signaling Peptides and Proteins ; KB Cells ; Lymphangiogenesis ; Mice ; Mice, Nude ; Mouth Floor ; Phosphotransferases ; Receptors, Vascular Endothelial Growth Factor ; Transplantation, Heterologous ; Transplants ; Tumor Burden ; Tyrosine ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factor Receptor-2 ; Vascular Endothelial Growth Factor Receptor-3

<b>OBJECTIVEb>To verify whether the extracellular domain of kinase domain region (KDR) has anti-angiogenesis activity in vivo.

<b>METHODSb>cDNA was cloned into adeno-associated virus (AAV) vector pSNAV and transfected to baby hamster kidney (BHK) cells. Recombinant AAV was obtained from the cell culture supernatant after adding helper virus. Recombinant AAV-infected human bladder cancer EJ cell line (EJ cells) were injected subcutaneously into Balb-c nude mice. Tumor specimens were removed from the mice, paraffin-embedded and sliced, then stained by immunohistochemistry. Microvessel density (MVD) was determined under a microscope.

<b>RESULTSb>The tumor volume developed by EJ cells transfected with the extracellular domain of KDR was significantly smaller (1.70 +/- 0.18 cm(3)) compared with that in the control (5.62 +/- 0.67 cm(3)) (P < 0.05), although tumor developed to be detectable on almost the same time (14.7 +/- 2.4 days vs 14.1 +/- 3.2 days). Further, MVD in the experimental group was lower than that in the control (41.3 +/- 4.8 vs 6.2 +/- 2.1, P < 0.05).

<b>CONCLUSIONb>The extracellular domain of KDR could be expressed in nude mouse bladder cancer tissue and inhibit tumor angiogenesis.

Animals ; Cloning, Molecular ; Cricetinae ; Dependovirus ; genetics ; Endothelial Growth Factors ; metabolism ; Female ; Genetic Therapy ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphokines ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; prevention & control ; Urinary Bladder Neoplasms ; blood supply ; therapy ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; Vascular Endothelial Growth Factors