Serum Vascular Endothelial Growth Factors A, C and D in Human Breast Tumors

Available evidence suggests that vascular endothelial growth factor (VEGF) a potent regulator of vasculogenesis and tumor angiogenesis may be a predictor of recurrence in breast cancer patients. We sought to determine whether VEGF serum levels (VEGF-A, VEGF-C and VEGF-D) in 377 patients with malignant and benign breast tumors differ and whether there is association between vascular growth factors, clinicopathologic features and prognosis. There was no significant difference in investigated circulating angiogenic markers between patients with malignant and non malignant lesions. We found strong correlation between VEGF-A and VEGF-D and between VEGF- C and VEGF-D. Besides serum VEGF-D levels and estrogen receptor (ER) expressions no other correlations between VEGF and clinicopathologic variables were observed. However, elevated VEGF-A and VEGF-C concentrations were associated with increased number of erythrocytes, leukocytes and platelets. In Cox model values of angiogenic serum markers and recognized prognostic markers in breast cancer, VEGF-C turned out as independent prognostic factor. Our study is the first analysis showing correlation between serum concentrations of three angiogenic factors: VEGF-A, VEGF-C, VEGF-D. Associations between angiogenic cytokines and number of blood cells may be due to release of VEGF from platelets and leucocytes. Prognostic role of VEGF is still uncertain, though VEGF-C has a potential to serve as a prognostic marker.     

VEGF-C,VEGF-D在乳腺癌中的表达及其与淋巴结转移及预后的关系

目的探讨VEGF-C和VEGF-D在乳腺癌组织中的表达以及与淋巴结转移、预后的相关性。方法采用SP免疫组化法检测78例乳腺癌癌组织和30例癌旁组织中VEGF-C,VEGF-D的表达水平,并完成所有患者的5年的随访资料。结果 78例乳腺癌癌组织中VEGF-C/D表达与癌旁组织相比差异有统计学意义(P<0.05);乳腺癌中VEGF-C表达与VEGF-D表达无相关性。VEGF-C,VEGF-D的表达水平与乳腺癌脉管内侵犯、淋巴结转移密切相关(P<0.05),但与年龄、肿瘤大小、TNM临床分期、ER、PR及Her-2的表达无关(P>0.05)。VEGF-C和VEGF-D的表达水平与乳腺癌的总生存期及无病生存期密切相关(P<0.05)。结论 VEGF-C,VEGF-D在乳腺癌组织中呈高表达;VEGF-C/D的表达水平与乳腺癌淋巴结转移能力和预后密切相关。     

Expression of vascular endothelial growth factor receptor-3 by lymphatic endothelial cells is associated with lymph node metastasis in prostate cancer.

PURPOSE: The molecular mechanisms underlying lymph node metastasis are poorly understood, despite the well-established clinical importance of lymph node status in many human cancers. Recently, vascular endothelial growth factor (VEGF)-C and VEGF-D have been implicated in the regulation of tumor lymphangiogenesis and enhancement of lymphatic invasion via activation of VEGF receptor-3. The purpose of this study was to determine the expression pattern of the VEGF-C/VEGF-D/VEGF receptor-3 axis in prostate cancer and its relationship with lymph node metastasis. EXPERIMENTAL DESIGN: The expression pattern of VEGF-C, VEGF-D, and VEGF receptor-3 in localized prostate cancer specimens (n = 37) was determined using immunohistochemistry. RESULTS: Widespread, heterogeneous staining for VEGF-C and VEGF-D was observed in all cancer specimens. Intensity of VEGF-C staining was lower in benign prostate epithelium than in adjacent carcinoma, whereas no difference between benign epithelium and carcinoma was observed for VEGF-D staining. VEGF receptor-3 immunostaining was detected in endothelial cells of lymphatic vessels in 18 of 37 tissue samples. The presence of VEGF receptor-3-positive vessels was associated with lymph node metastasis (P = 0.0002), Gleason grade (P < 0.0001), extracapsular extension (P = 0.0382), and surgical margin status (P = 0.0069). In addition, VEGF receptor-3 staining highlighted lymphatic invasion by VEGF-C-positive/VEGF-D-positive carcinoma cells. CONCLUSIONS: Together, these results suggest that paracrine activation of lymphatic endothelial cell VEGF receptor-3 by VEGF-C and/or VEGF-D may be involved in lymphatic metastasis. Thus the VEGF-C/VEGF-D/VEGF receptor-3 signaling pathway may provide a target for antilymphangiogenic therapy in prostate cancer.     

Pre-operative plasma levels of vascular endothelial growth factor A, C and D in patients with colorectal cancer

AimsVascular endothelial growth factor (VEGF)-C and VEGF-D are angiogenic and lymphangiogenic members of the VEGF family of growth factors. Increased VEGF-C or VEGF-D expression in human tumours may be associated with lymph-node metastasis and lymphatic invasion. Circulating plasma levels of VEGF-A, VEGF-C and VEGF-D were measured in patients with colorectal cancer, and assessed for their usefulness as a diagnostic tool for determining lymph-node metastasis.Materials and methodsOne hundred and twenty patients with colorectal cancer and 50 healthy control patients were included in the study. Plasma growth-factor levels were assessed by enzyme-linked immunosorbent assays.ResultsNo significant differences in plasma VEGF-C or VEGF-D levels were seen between patients subgrouped by clinicopathological variables. In particular, there were no differences in median plasma VEGF-C or VEGF-D level in patients with and without lymph-node involvement (VEGF-C: 11.2 U/ml [range, 4.9–51.9] vs 9.9 U/ml [4.4–93.4 U/ml]; P = 0.90; VEGF-D: 335 pg/ml [113–1102] vs 316.5 pg/ml [0–1343]; P = 0.68).ConclusionsCirculating plasma levels of VEGF-C and VEGF-D do not allow pre-operative identification of lymph-node status in patients with colorectal cancer.     

Clinical significance of VEGF-A, -C and -D expression in esophageal malignancies

Vascular endothelial growth factors (VEGF)-A, -C and -D are members of the proangiogenic VEGF family of glycoproteins. VEGF-A is known to be the most important angiogenic factor under physiological and pathological conditions, while VEGF-C and VEGF-D are implicated in the development and sprouting of lymphatic vessels, so called lymphangiogenesis. Local tumor progression, lymph node metastases and hematogenous tumor spread are important prognostic factors for esophageal carcinoma (EC), one of the most lethal malignancies throughout the world. We found solid evidence in the literature that VEGF expression contributes to tumor angiogenesis, tumor progression and lymph node metastasis in esophageal squamous cell carcinoma (SCC), and many authors could show a prognostic value for VEGF-assessment. In adenocarcinoma (AC) of the esophagus angiogenic properties are acquired in early stages, particularly in precancerous lesions like Barrett's dysplasia. However, VEGF expression fails to give prognostic information in AC of the esophagus. VEGF-C and -D were detected in SCC and dysplastic lesions, but not in normal mucosa of the esophagus. VEGF-C expression might be associated with lymphatic tumor invasion, lymph node metastases and advanced disease in esophageal SCC and AC. Therapeutic interference with VEGF signaling may prove to be a promising way of anti-angiogenic co-treatment in esophageal carcinoma. However, concrete clinical data are still pending.     

Vascular endothelial growth factor C and vascular endothelial growth factor receptor 2 are related closely to the prognosis of patients with ovarian carcinoma

BACKGROUND: The vascular endothelial growth factor (VEGF) family and VEGF receptors (VEGFR) play an essential role in the angiogenesis of both pathologic and nonpathologic conditions. However, the prognostic significance of VEGF and VEGFR expression in ovarian carcinoma is unclear. METHODS: The tissue expression levels of VEGF-A, VEGF-C, VEGFR-2, and VEGFR-3 in 80 specimens of ovarian carcinoma were examined immnohistochemically. The results obtained were analyzed clinicopathologically. RESULTS: VEGF-A, VEGF-C, VEGFR-2, and VEGFR-3 were expressed both in tumor cells and in adjacent endothelial cells of blood and lymph vessels. The tissue expressions of VEGF-C and VEGFR-2 were correlated significantly with tumor extension, including peritoneal metastases outside the pelvic cavity (P = 0.0010 and P = 0.0008, respectively), lymph node metastases (P = 0.0030 and P = 0.0018, respectively), and positive ascitic cytology (P = 0.025 and P = 0.0016, respectively). Conversely, there was no significant correlation between VEGF-A and VEGFR-3 expression and clinicopathologic features of ovarian carcinoma. Logistic regression analysis revealed that the expressions of VEGF-C and VEGFR-2 also were independent risk factors for peritoneal and lymph node metastases. Survival curves determined by the Kaplan-Meier method and in univariate analysis demonstrated that high expression levels of VEGF-C and VEGFR-2 were associated with the 5-year survival rate. In multivariate analysis, high expression levels of VEGF-C and VEGFR-2 emerged as independent indicators for disease-specific survival. CONCLUSIONS: High tissue expression of VEGF-C and VEGFR-2 reflects the aggressiveness of the spread of tumor in ovarian carcinoma. Thus, both have predictive value for identifying high-risk patients who have a poor prognosis.     

Clinicopathological and prognostic significance of vascular endothelial growth factors (VEGF)-C and -D and VEGF receptor 3 in invasive breast carcinoma.

AIMS: Vascular endothelial growth factors C and D (VEGF-C and VEGF-D) play a major role in lymphangiogenesis and activate VEGF receptor 3 (VEGFR-3). Our purpose was to study the clinicopathologic and clinical value of VEGF-C, VEGF-D and VEGFR-3 in invasive breast carcinoma. MATERIAL AND METHODS: Immunohistochemistry was performed in paraffin-embedded tissue specimens from 177 invasive breast carcinomas to detect the proteins VEGF-C, VEGF-D, VEGFR-3, p53, Ki67, c-erbB-2, topoII alpha and ER/PR. The results were statistically processed. RESULTS: VEGF-C, VEGF-D and VEGFR-3 were found to be predominantly expressed in the cytoplasm of the malignant cells. VEGF-C occasionally showed a submembranous intensification. VEGF-D and VEGFR-3 were also immunodetected in the nuclei of the malignant cells. Nuclear VEGF-D was positively correlated to p53, Ki67 and topoII alpha proteins' expression (p=0.003, p=0.009 and p=0.017 respectively) and nuclear VEGFR-3 to topoII alpha (p=0.034). Cytoplasmic expression of VEGF-C and its submembranous intensification were found to be independent indicators of patients' overall and disease-free survival, respectively (p=0.003 and p=0.044 respectively). The group with high expression of both cytoplasmic VEGF-C and stromal VEGFR-3 showed poor overall survival (p=0.024) and the group with both submembranous VEGF-C and stromal VEGFR-3 immunostaining showed poor both disease-free and overall survival (p=0.012 and p=0.038 respectively). CONCLUSION: VEGF-D and VEGFR-3 seem to exert proliferative activity in invasive breast carcinomas. VEGF-C was found to be an independent indicator of patient's poor prognosis and the simultaneous expression of tumor VEGF-C and stromal VEGFR-3 yielded additional prognostic information.     

VEGF、VEGF-C和VEGF-D在甲状腺乳头状癌组织中的表达及意义

背景与目的:研究表明血管内皮生长因子C和D(vascularendothelialgrowthfactor-Cand-D,VEGF-C,VEGF-D)与淋巴管的生成有关。局部淋巴结转移是甲状腺乳头状癌的特征,本研究的目是探讨血管内皮生长因子及其C和D(VEGF、VEGF-C和VEGF-D)蛋白在甲状腺乳头状癌的表达及其意义。方法:应用免疫组织化学SP法检测115例甲状腺乳头状癌和20例结节性甲状腺肿中VEGF、VEGF-C和VEGF-D蛋白的表达。结果:甲状腺乳头状癌VEGF、VEGF-C和VEGF-D的阳性率分别为79.1%、87.0%和72.2%。结节性甲状腺肿组织中VEGF、VEGF-C和VEGF-D的阳性率分别为30.0%、15.0%和20.0%。甲状腺乳头状癌VEGF、VEGF-C和VEGF-D的阳性率较结节性甲状腺肿明显增高。VEGF的表达与甲状腺癌的大小有关。VEGF、VEGF-C和VEGF-D在淋巴结转移组和非转移组的阳性率分别为84.7%、93.2%、83.1%和73.2%、80.4%、60.7%。VEGF-C和VEGF-D的阳性表达与甲状腺乳头状癌的淋巴结转移显著相关(P<0.05;P<0.01)。结论:VEGF与甲状腺乳头状癌的生长有关。VEGF-C和VEGF-D与淋巴结转移密切相关,提示甲状腺乳头状癌标本VEGF-C和VEGF-D的检测可作为预测肿瘤转移的指标之一。     

Vascular endothelial growth factors C and D and lymphangiogenesis in gastrointestinal tract malignancy

Vascular endothelial growth factor-C (VEGF-C) and VEGF-D are members of the VEGF family of cytokines and have angiogenic and lymphangiogenic actions. In gastric adenocarcinoma, VEGF-C mRNA and tissue protein expression correlate with lymphatic invasion, lymph node metastasis and in some reports, venous invasion and reduced 5-year survival. Patients with gastric adenocarcinomas containing high levels of VEGF-C expression have significantly reduced 5-year survival rates, and VEGF-C expression is an independent prognostic risk factor for death. The role of VEGF-C in oesophageal squamous and colorectal cancer and VEGF-D in colorectal cancer is not clear, with conflicting reports in the published literature. In order to exploit potential therapeutic applications, further research is necessary to define the precise roles of these cytokines in health and disease.     

MIA-dependent angiogenesis and lymphangiogenesis are closely associated with progression,nodal metastasis and poor prognosis in tongue squamous cell carcinoma

We examined the role of angiogenesis/lymphangiogenesis and the relationship between melanoma inhibitory activity (MIA) and angiogenesis or lymphangiogenesis in oral squamous cell carcinoma (OSCC). One hundred and one formalin-fixed, paraffin-embedded specimens of primary OSCC were evaluated for microvessel density (MVD), lymphovessel density (LVD), expression of vascular endothelial growth factor (VEGF), VEGF-C, VEGF-D and MIA. Fresh frozen 18 samples of primary OSCC were further examined for the expression of VEGF, VEGF-C, VEGF-D and MIA protein by enzyme-linked immunosorbent assay (ELISA). In in vitro analysis, we studied the change of VEGF, VEGF-C and VEGF-D expression after MIA siRNA treatment. Higher MVD, LVD and VEGF expression levels were closely associated with tumour progression, nodal metastasis and poor prognosis. Expression levels of VEGF-C and VEGF-D were only related with nodal metastasis. MIA expression was significantly associated with MVD, LVD, VEGF, VEGF-C and VEGF-D expression by immunohistochemistry and ELISA assay. VEGF, VEGF-C, VEGF-D and MIA expression levels of metastatic tongue cancer HSC-3 cells were higher than those with no metastatic HSC-4 cells, and VEGF, VEGF-C and VEGF-D expression levels were decreased by MIA siRNA treatment in both cells. MIA-dependent angiogenesis/lymphangiogenesis might be a useful therapeutic target in progressive and metastatic OSCC.     

The association between vascular endothelial growth factor-C, its corresponding receptor, VEGFR-3, and prognosis in primary breast cancer: a study with 193 cases

Lymphangiogenesis plays an important role in several normal and pathological conditions, such as wound healing, pathogen infection, inflammation or the metastasis formation of endothelial malignancies. Vascular endothelial growth factor-C (VEGF-C) and VEGF-D are important and specific regulatory factors for lymphatic endothelial proliferation and lymphangiogenesis. Both growth factors mediate their biological activity mainly by VEGF receptor-3 (VEGFR-3, Flt-4). In this study, we measured intratumoral levels of VEGF-C and VEGFR-3 through enzyme-linked immunosorbent assay (ELISA) in 193 primary breast cancer tissues and examined their prognostic values. A significant correlation was found between the VEGF-C and VEGFR-3 protein levels. High VEGF-C levels were associated with low-grade tumors and a smaller size. Univariate analysis showed that high VEGF-C was significantly associated with a favourable prognosis for disease-free survival (DFS) and overall survival (OS). No significant prognostic value of VEGFR-3 was detected. Multivariate analysis confirmed the independent prognostic value of VEGF-C. The intratumoral VEGF-C level is a significant prognostic indicator of primary breast cancer. An investigation of the mechanisms of VEGF-C protein processing in human cancer tissue should be carried out in the future.     

Pilot study to relate clinical outcome in pancreatic carcinoma and angiogenic plasma factors/circulating mature/progenitor endothelial cells: Preliminary results

Circulating endothelial cells (CEC) and bone marrow-derived endothelial progenitors (ECP) play important roles in tumor growth and have been proposed as non-invasive markers of angiogenesis. However, CEC and ECP levels have not been investigated in pancreatic carcinoma patients. Using four-color flow cytometry procedures, we evaluated the count of resting (rCEC) and activated (aCEC) endothelial cells and ECP in the peripheral blood of pancreatic carcinoma patients before and after chemotherapy, consisting of gemcitabine (GEM) alone or in combination with oxaliplatin (OX), or with 5-fluorouracil (5-FU). We also correlated CEC and ECP levels with plasma levels of relevant angiogenic factors, such as vascular endothelial growth factor (VEGF)-A, VEGF-D, angiopoietin (Angio)-1, and chemokine C-X-C motif ligand (CXCL)12, measured by ELISA, and with clinical features of pancreatic cancer. The aCEC, rCEC, ECP, and VEGF-A plasma levels were significantly higher in locally-advanced and metastatic patients than controls. Both ECP and VEGF-A levels correlated positively with disease stage and inversely with patient's overall survival. Measurements after the treatment course showed that VEGF-A plasma concentrations and ECP counts had decreased significantly. In particular, VEGF-A and rCEC were significantly down after treatment with GEM alone or in combination with OX. No significant differences in terms of circulating angiogenic factor or endothelial cell subtype levels were found between responders (patients entering partial remission or with stable disease) and non-responders (patients with progressive disease). The study provides insights into angiogenesis mechanisms in pancreatic carcinoma, for which anti-angiogenic targeting of VEGF-A and ECP could be of interest.     

ErbB2 overexpression in human breast carcinoma is correlated with p21Cip1 up-regulation and tyrosine-15 hyperphosphorylation of p34Cdc2: poor responsiveness to chemotherapy with cyclophoshamide methotrexate,and 5-fluorouracil is associated with Erb2 overexpression and with p21Cip1 overexpression

BACKGROUND: Clinical investigations have shown that in patients with breast carcinoma, tumors that overexpress the erb-B2 gene are less responsive to certain chemotherapy regimens compared with tumors that express low levels of ErbB2, suggesting that ErbB2 overexpression may be used as a marker for poor response to chemotherapy in patients with breast carcinoma. The combination of cyclophosphamide, methotrexate, and 5-fluorouracil (CMF) is one of the most widely used chemotherapy regimens in patients with breast carcinoma. Patients who have ErbB2-overexpressing breast carcinomas have poorer responses to CMF compared with patients who have breast carcinomas with low ErbB2 expression. ErbB2-overexpressing breast tumor cells are resistant to taxol-induced apoptotic cell death. The underlying molecular mechanism is that ErbB2 inhibits p34(Cdc2) activation, which is required for taxol-induced apoptosis, by up-regulating p21(Cip1) and by hyperphosphorylating p34(Cdc2) on tyrosine-15. However, the relation between ErbB2, p21(Cip1), and p34(Cdc2) in patients with breast carcinoma remains elusive. The contribution of these molecular alterations to ErbB2-mediated CMF resistance has not been examined. METHODS: Formalin-fixed, paraffin-embedded, 5 microm thick tissue sections from 107 patients with invasive breast carcinoma were immunostained using specific antibodies against ErbB2, p21(Cip1), and phosphorylated tyrosine (Tyr)-15 of p34(Cdc2). Ninety-four of 107 patients were treated with the CMF regimen. In situ hybridization of p21(Cip1)mRNA also was performed in 20 of the sections described above. ErbB2 expression levels, p21(Cip1) expression levels, and phosphorylation status on Tyr15 of p34(Cdc2) were analyzed for correlations with clinicopathologic parameters for the 107 patients and for correlations with disease-free survival (DFS) in the 94 patients who were treated with the CMF regimen. RESULTS: Among 94 patients with breast carcinoma who were treated with CMF, it was found that ErbB2 overexpression was associated significantly with poor DFS (P < 0.01). Patients who had higher p21(Cip1) expression had worse DFS compared with patients who had low p21(Cip1) expression (P = 0.02). However, no significant correlation was found between p34(Cdc2)-Tyr15 phosphorylation and DFS (P > 0.05). It is noteworthy that p21(Cip1) expression and p34(Cdc2)-Tyr15 phosphorylation were correlated significantly and positively with ErbB2 expression (P < 0.01). CONCLUSIONS: The current study suggests that p21(Cip1) expression, but not p34(Cdc2)-Tyr15 phosphorylation, may play a role in ErbB2-mediated CMF resistance, which may contribute to the poor survival of patients with ErbB2-overexpressing breast carcinomas who were treated on the CMF regimen. In addition, ErbB2 overexpression was correlated with p21(Cip1) up-regulation and with increased p34(Cdc2)-Tyr15 phosphorylation in breast tumors.