PURPOSE: Animal models show a strong relationship between lymphangiogenesis and lymph node metastasis. However, the clinical significance of lymphangiogenesis in patients with colorectal cancer (CRC) remains uncertain. This study aimed to evaluate the association between c-Met and lymphangiogenic factors and to elucidate the prognostic significance of c-Met in patients with CRC. METHODS: A total of 379 tissue samples were obtained from surgically resected specimens from patients with CRC at Soonchunhyang University Cheonan Hospital between January 2002 and December 2010. The expressions of c-Met, vascular endothelial growth factor (VEGF)-C, VEGF-D, VEGF receptor (VEGFR)-3, and podoplanin were examined using immunohistochemistry. The expression of c-Met and clinical factors were analyzed. RESULTS: Of the 379 tissues, 301 (79.4%) had c-Met expression. High expression of c-Met in tumor cells was significantly associated with high expression of VEGF-C (P < 0.001) and VEGFR-3 (P = 0.001). However, no statistically significant association with podoplanin (P = 0.587) or VEGF-D (P = 0.096) was found. Of the 103 evaluable patients, expression of c-Met in tumor cells was significantly associated with advanced clinical stage (P = 0.020), positive lymph node status (P = 0.038), and high expression of VEGF-C (P = 0.020). However, no statistically significant association with podoplanin (P = 0.518), VEGFR-3 (P = 0.085), VEGF-D (P = 0.203), or overall survival (P = 0.360) was found. CONCLUSION: Our results provide indirect evidence for an association and possible regulatory link of c-Met with the lymphangiogenic markers, but c-Met expression in patients with CRC is not a prognostic indicator for overall survival.
Chungcheongnam-do ; Colorectal Neoplasms* ; Humans ; Immunohistochemistry ; Lymph Nodes ; Lymphangiogenesis ; Models, Animal ; Neoplasm Metastasis ; Receptors, Vascular Endothelial Growth Factor ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factor D ; Vascular Endothelial Growth Factor Receptor-3

Vascular endothelial growth factor (VEGF) exerts its biological functions by its specific VEGF receptors (VEGFR), which includes VEGFR-1, VEGFR-2, VEGFR-3, neuropilin-1, and neuropilin-2. These VEGFR distributes in endothelial cells, and are also expressed in normal skin, inflammatory skin diseases, and skin cancers. The VEGF-VEGFR signaling pathway may be a new key target in the management of the skin diseases.
Animals ; Humans ; Receptors, Vascular Endothelial Growth Factor ; biosynthesis ; Signal Transduction ; Skin Diseases ; metabolism ; Vascular Endothelial Growth Factor A ; physiology

PURPOSE: Nodal metastasis is the main prognostic factor in the patients with oral squamous cell carcinoma (OSCC). We investigated the association between tumor-associated lymphatics and OSCC characteristics. METHODS: Thirty-four specimens were used for the immunohistochemical staining with the antibody for vascular endothelial growth factor (VEGF)-C, VEGF-D, VEGF receptor (VEGFR)-3, phosphorylated VEGFR-3, D2-40, and matrix metallproteinases (MMPs). We observed the distribution of the lymphangiogenic factors and quantified the degree of expression. We determined lymphatic vessel density (LVD) and lymphatic vessel dilatation with D2-40 immunostaining. We assessed the association of LVD or lymphatic vessel dilatation with tumor progression or tumor differentiation. RESULTS: OSCC cells expressed lymphangiogenic ligands. Lymphangiogenic receptor, VEGFR-3, was expressed and activated in some tumor cells as well as in tumor-associated endothelial cells. LVD was not associated with tumor size or nodal status, but lymphatic vessel dilatation was higher in tumors with nodal metastasis, and also higher in poorly differentiated tumors. In stromal area of OSCC, MMP-1 and MMP-10 were up-regulated and the basement membrane of tumor-associated endothelial cells was destroyed by these collagenases. CONCLUSION: In the primary tumors with nodal metastasis, especially in poorly differentiated OSCC, tumor cells invaded the dilated lymphatic vessels via ruptured sites. MMP-1 and MMP-10 are important in the lysis of the glycocalyx inside the tumor-associated lymphatic endothelial cells.
Basement Membrane ; Carcinoma, Squamous Cell* ; Collagenases ; Dilatation ; Endothelial Cells ; Glycocalyx ; Humans ; Ligands ; Lymphatic Vessels ; Neoplasm Metastasis* ; Receptors, Vascular Endothelial Growth Factor ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor D ; Vascular Endothelial Growth Factor Receptor-3

Here, we investigated the role of zerumbone, a natural cyclic sesquiterpene of Zingiber zerumbet Smith, on angiogenesis using human umbilical vein endothelial cells (HUVECs). Zerumbone inhibited HUVECs proliferation, migration and tubule formation, as well as angiogenic activity by rat aorta explants. In particular, zerumbone inhibited phosphorylation of vascular endothelial growth factor receptor-2 and fibroblast growth factor receptor-1, which are key regulators of endothelial cell function and angiogenesis. In vivo matrigel plug assay in mice demonstrated significant decrease in vascularization and hemoglobin content in the plugs from zerumbone-treated mice, compared with control mice. Overall, these results suggest that zerumbone inhibits various attributes of angiogenesis, which might contribute to its reported antitumor effects.
Animals ; Aorta ; Endothelial Cells ; Fibroblast Growth Factors ; Ginger* ; Human Umbilical Vein Endothelial Cells ; Mice ; Phosphorylation ; Rats ; Receptors, Vascular Endothelial Growth Factor ; Vascular Endothelial Growth Factor Receptor-2

OBJECTIVE: To Investigate the expression of vascular endothelial growth factor receptor-1, -2, and -3 (VEGFR-1, -2, and -3) mRNA in the eutopic and ectopic tissues in women with endometriosis. METHODS: At the time of laparoscopy or laparotomy for endometriosis, infertility or other benign gynecologic conditions, a biopsy specimen was taken from the endometrial cavity and a peritoneal endometriotic implant simultaneously whenever appropriate. For control samples, endometrial tissues were obtained from women without visual evidence of endomtriosis. We employed real time quantitative RT-PCR to quantify VEGFR gene expression of these tissues. Comparisions between each group were made using ANOVA (analysis of variance) and Kruskal-Wallis test and statistical significance was defined as p<0.05. RESULTS: The expression of VEGFR-1, -2, and, -3 mRNA was significantly higher in both eutopic and ectopic endometrial tissues of women with endometriosis than in control endometrial tissues during both proliferative and mid-secretory phase. In ectopic endometrial tissue, VEGFR-1 mRNA expression was significantly increased during the mid-secretory phase compared to the proliferative phase. There was a marked increase especially in VEGFR-3 mRNA expression in ectopic endometriotic lesions during the proliferative phase but its expression decreased during the mid-secretory phase. CONCLUSION: mRNA for VEGFR-1, -2, and -3 in an endometriotic lesions might be differentially expressed and their expression appears to be associated with the development of endometriosis.
Biopsy ; Choristoma ; Endometriosis* ; Female ; Gene Expression ; Humans ; Infertility ; Laparoscopy ; Laparotomy ; Receptors, Vascular Endothelial Growth Factor* ; RNA, Messenger ; Vascular Endothelial Growth Factor A* ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-3

BACKGROUND: Colorectal cancer (CRC) is one of the most common malignant neoplasms and is a leading cause of mortality worldwide. Metastasis to the liver is a frequent event in patients with CRC. An essential step in the metastatic cascade is angiogenesis. METHODS: This study included 45 patients who underwent a partial colectomy with hepatic resection for CRC with hepatic metastases. Immunohistochemistry was performed using vascular endothelial growth factor (VEGF)-A, VEGF receptor (VEGFR)-1, VEGFR-2, and CD34 antibodies to examine the relationship between CRC with liver metastases and angiogenesis. RESULTS: CRC showed significantly stronger expression of VEGF-A, VEGFR-1, and VEGFR-2 than liver metastases (p < 0.05). Microvessel density was also higher in CRC than in liver metastases (p < 0.05). CONCLUSIONS: Compared with previous studies, we found a higher expression of VEGF-A, VEGFR-1, VEGFR-2, and microvessel density in CRC than in liver metastases, which could be ascribed to a difference in vessel distribution and blood supply in each organ. Given its profuse blood supply and distinct cell populations, the liver might provide a rich milieu for tumor cell growth with less expression of angiogenesis-inducing agents.
Antibodies ; Colectomy ; Colorectal Neoplasms ; Glycosaminoglycans ; Humans ; Immunohistochemistry ; Liver ; Microvessels ; Neoplasm Metastasis ; Receptors, Vascular Endothelial Growth Factor ; Thymus Gland ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-2 ; World Health Organization

Vascular endothelial growth factor (VEGF) exerts its biological functions by its specific VEGF receptors (VEGFRs), which includes VEGFR-1, VEGFR-2, VEGFR-3, neuropilin-1 and neuropilin-2. These VEGF receptors not only distribute in endothelial cells, but also in epidermal keratinocytes. VEGFRs may play a significant role in pathogenesis of the epidermal neoplasm and the VEGF-VEGFR signaling pathway may be a novel therapy target for neoplasm derived from epidermis.
Animals ; Epidermis ; metabolism ; Humans ; Neoplasms ; metabolism ; Neuropilins ; genetics ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-3 ; genetics ; metabolism

This study was to evaluate the expression of vascular endothelial growth factor receptors (VEGFRs) in tumor and stromal cells of tougue squamous cell carcinoma (SCC). We also wanted to characterize the differences, from the angiogenic aspect, between cancer-associated stromal cells and non-malignant stromal cells. Paraffin-embedded tumor specimens from eleven patients with tongue SCCs were studied. Immunohistochemical staining for VEGFR-1,-2, and -3 was performed on the tumor cells, stromal fibroblasts and tumor-associated macrophages of the specimens. The expression of all 3 receptors was detected in the tumor cells themselves of the biopsy specimens. All 3 receptors were also expressed on stromal cells, except that VEGFR-2 was not expressed in stromal fibroblasts. In radical excision specimens, the staining intensity for VEGFR-1, -2 in the tumor cells and VEGFR-1,-3 in the tumor-associated macrophages was significantly lower than that in the biopsy specimens (P < 0.05). By using the general marker of fibroblast and macrophage, 5B5 and CD68, respectively, we performed double immunofluorescence staining for 5B5 and each VEGFR in the stromal fibroblasts and for CD68 and each VEGFR in the tumor-associated macrophages of the radical excision specimens. We used 4 cases of fibroma and 4 cases of chronic inflammation tissue as the controls. It was found that only each marker was expressed in the control group, however, 5B5/VEGFR-1 and 5B5/VEGFR-3 in the stromal fibroblasts, and CD68/VEGFR-1 and CD68/VEGFR-3 in the tumor-associated macrophages were double stained in the radical excision specimens. Although our study used small number of specimens, the results of our study showed that in tongue SCC, in association with the angiogenesis, the stromal cells showed the activated phenotype and this was different from the nonmalignant stromal cells.
Biopsy ; Carcinoma, Squamous Cell* ; Fibroblasts ; Fibroma ; Fluorescent Antibody Technique ; Humans ; Inflammation ; Macrophages ; Phenotype ; Receptors, Vascular Endothelial Growth Factor* ; Stromal Cells* ; Tongue* ; Vascular Endothelial Growth Factor A* ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-2

OBJECTIVE: To investigate the expressions of stromal cell-derived factor (CXCL12), stromal cell-derived factor receptor (CXCR4), vascular endothelial growth factor (VEGF) and microvessel density (MVD) in bone marrow microsputum of patients with multiple myeloma (MM) and their correlation with the prognosis. METHODS: The expressions of CXCL12, CXCR4, VEGF and MVD in bone marrow microtubules of 57 newly diagnosed MM patients and 26 normal bone marrow samples were detected by immunohistochemistry. The rank sum test was used to compare the differences between the two groups. The clinical data of the patients were collected to analyze the correlation between the indicators of the MM group and the prognosis. RESULTS: The expressions of CXCL12, CXCR4, VEGF and MVD in the bone marrow biopsy of the patients in MM group were significantly higher than those in the normal control group (P＜0.05). The expressions levels of CXCL12, CXCR4, VEGF and MVD were in the bone marrow of the patients in MM group were correlated with the ISS stage, risk stratification and the proportion of plasma cells in the bone marrow (P＜0.05). Univariate analysis showed that age, ISS stage, risk stratification, plasma cell ratio, expressions of CXCL12, CXCR4, VEGF, and MVD associated with the prognosis of patients with MM (P＜0.05). Multivariate analysis found that expressions of CXCR4, VEGF, MVD, age, and plasma cell ratio were independent prognostic factors. CONCLUSION The expressions of CXCL12, CXCR4, VEGF and MVD are increase in the bone marrow of patients with multiple myeloma, and their expressions levels are associate with the occurrence and development of multiple myeloma, and their high expression may indicate a poor prognosis.
Chemokine CXCL12 ; Humans ; Multiple Myeloma ; Neovascularization, Pathologic ; Patients ; Prognosis ; Receptors, CXCR4 ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

BACKGROUND: Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis, including stimulating the proliferation and migration of vascular smooth muscle cells (VSMCs). It has been known that diabetes is associated with accelerated cellular proliferation via VEGF, as compared to that under a normal glucose concentration. We investigated the effects of selective blockade of a VEGF receptor by using anti-Flt-1 peptide on the formation and hyperplasia of the neointima in balloon injured-carotid arteries of OLETF rats and also on the in vitro VSMCs' migration under high glucose conditions. MATERIAL AND METHOD: The balloon-injury method was employed to induce neointima formation by VEGF. For 14 days beginning 2 days before the ballon injury, placebo or vascular endothelial growth factor receptor-1 (VEGFR-1) specific peptide (anti-Flt-1 peptide), was injected at a dose of 0.5 mg/kg daily into the OLETF rats. At 14 days after balloon injury, the neointimal proliferation and vascular luminal stenosis were measured, and cellular proliferation was assessed by counting the proliferative cell nuclear antigen (PCNA) stained cells. To analyze the effect of VEGF and anti-Flt-1 peptide on the migration of VSMCs under a high glucose condition, transwell assay with a matrigel filter was performed. And finally, to determine the underlying mechanism of the effect of anti-Flt-1 peptide on the VEGF-induced VSMC migration in vitro, the expression of matrix metalloproteinase (MMP) was observed by performing reverse transcription-polymerase chain reaction (RT-PCR). RESULT: Both the neointimal area and luminal stenosis associated with neointimal proliferation were significantly decreased in the anti-Flt-1 peptide injected rats, (0.15+/-0.04 mm2 and 36.03+/-3.78% compared to 0.24+/-0.03 mm2 and 61.85+/-5.11%, respectively, in the placebo-injected rats (p<0.01, respectively). The ratio of PCNA(+) cells to the entire neointimal cells was also significantly decreased from 52.82+/-4.20% to 38.11+/-6.89% by the injected anti-Flt-1 peptide (p<0.05). On the VSMC migration assay, anti-Flt-1 peptide significantly reduced the VEGF-induced VMSC migration by about 40% (p<0.01). Consistent with the effect of anti-Flt-1 peptide on VSMC migration, it also obviously attenuated the induction of the MMP-3 and MMP-9 mRNA expressions via VEGF in the VSMCs. CONCLUSION: Anti-Flt-1 peptide inhibits the formation and hyperplasia of the neointima in a balloon-injured carotid artery model of OLETF rats. Anti-Flt-1 peptide also inhibits the VSMCs' migration and the expressions of MMP-3 and MMP-9 mRNA induced by VEGF under a high glucose condition.
Animals ; Arteries ; Carotid Arteries ; Cell Proliferation ; Constriction, Pathologic ; Endothelial Growth Factors* ; Glucose ; Hyperplasia ; Muscle, Smooth, Vascular* ; Neointima* ; Phenobarbital ; Rats* ; Rats, Inbred OLETF ; Receptors, Vascular Endothelial Growth Factor ; RNA, Messenger ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-1