Vascular endothelial growth factor-C expression in human prostatic carcinoma and its relationship to lymph node metastasis.

Lymph node dissemination is a major prognostic factor in human cancer. However, the molecular mechanisms underlying lymph node metastasis are poorly understood. Recently, vascular endothelial growth factor-C (VEGF-C) was identified as a ligand for VEGF receptor-3 (VEGFR-3/Flt-4) and the expression of VEGFR-3 was found to be highly restricted to the lymphatic endothelial cells. In this report, we investigated the expression of VEGF-C and VEGFR-3 in human prostatic carcinoma tissue by using in situ hybridization and immunohistochemical staining respectively. Expression of VEGF-C mRNA in prostatic carcinoma was significantly higher in lymph node-positive group than in lymph node-negative group. In addition, the number of VEGFR-3-positive vessels was increased in stroma surrounding VEGF-C-positive prostatic carcinoma cells. These results suggest that the expression of VEGF-C in prostatic carcinoma cells is implicated in the lymph node metastasis.     

血管内皮生长因子D及其受体3在乳腺癌淋巴转移中作用的研究进展

乳腺癌是女性最常见的恶性肿瘤。乳腺癌淋巴转移与患者的预后密切相关。最近研究表明淋巴管生成可能会主动促进淋巴转移的发生,而血管内皮生长因子家族的部分成员在这一过程中发挥了重要作用,如血管内皮生长因子C、血管内皮生长因子D及其受体3等。但是,对乳腺癌中血管内皮生长因子D的作用及其预后价值、血管内皮生长因子受体3与乳腺癌淋巴管生成的关系等问题仍有争议。本文对近年国内外有关血管内皮生长因子D及其受体3在乳腺癌淋巴转移中作用的研究进展作一综述。     

Vascular endothelial growth factor-D is an independent prognostic factor in epithelial ovarian carcinoma

We assessed the presence of vascular endothelial growth factor (VEGF)-C, VEGF-D and their receptor VEGFR-3 by immunohistochemistry in 59 epithelial ovarian carcinomas, 11 borderline tumours and 20 benign cystadenomas. VEGF-C and VEGF-D were generally expressed in tumour cells and also in endothelia adjacent to tumour nests which showed a strong staining for them. VEGFR-3 was expressed in lymphatic and vascular endothelial cells adjacent to tumour nests. Immunoreactivity was significantly more frequent as lesions progressed from a benign tumour to advanced carcinoma. A strong correlation was found between VEGF-C and VEGF-D detected in carcinoma and VEGFR-3 detected in neighbouring endothelial cells. Increased expression of VEGF-C, VEGF-D and VEGFR-3 was significantly associated with lymph node metastasis and peritoneal metastasis outside the pelvis. There was a significant correlation between the high levels of VEGF-C and VEGF-D proteins, and poor survival. The presence of VEGF-D was an independent prognostic indicator by multivariate analysis. We conclude that VEGF-C, VEGF-D and VEGFR-3 play an important role in lymphatic spread and intraperitoneal tumour development in ovarian carcinoma. Since VEGF-D was found to be an independent predictor of poor outcome, its measurement, together with other prognostic markers may improve prospective identification of patients with a poor prognosis.     

Expression analysis of vascular endothelial growth factors and their relationships to lymph node metastasis in human colorectal cancer

This study was undertaken to determine whether expressions of the vascular endothelial growth factor (VEGF) family (VEGF-A, VEGF-B, VEGF-C, and VEGF-D) are correlated with clinicopathological parameters, with particular reference to lymph node metastasis in colorectal cancer. Total RNA was isolated from 82 surgical specimens of colorectal cancer and matched to normal mucosa with (n = 41) or without (n = 41) lymph node metastasis. The mRNA expression of each VEGF family member was quantified by real-time quantitative (RTQ) RT-PCR assay. VEGF-B and VEGF-C mRNA were significantly higher both in the tumors with lymph node metastasis (p = 0.027 and p = 0.024, respectively) and in tumors with lymphatic invasion (p = 0.042 and p = 0.005, respectively). In contrast, VEGF-D mRNA was down-regulated in tumors with lymphatic involvement (p = 0.047). Among the other clinicopathological factors, we noted that VEGF-A mRNA was higher in tumors with liver metastasis than in those without (p = 0.018) and was higher in tumors with venous invasion than in those without (p = 0.007). The results of this study demonstrate that high levels of VEGF-B, C and low levels of VEGF-D mRNA expression are associated with lymph node metastasis and lymphatic involvement. These results suggest that a balance among VEGF-B, VEGF-C, and VEGF-D might contribute to the lymphangiogenic process and metastasis in colorectal cancer.     

Expression of vascular endothelial growth factor (VEGF)-C and VEGF-D in early gastric carcinoma: correlation with clinicopathological parameters

In this study, we investigated whether expression of vascular endothelial growth factor (VEGF)-C and/or VEGF-D correlates with clinicopathological features of human gastric carcinoma. We immunohistochemically examined the expression of VEGF-C and VEGF-D in 140 archival surgical specimens of submucosally invasive gastric carcinoma. Of these specimens, 32 (22.9%) and 12 (8.6%) showed intense VEGF-C and VEGF-D immunoreactivity in cancer cells, respectively. VEGF-C immunoreactivity was associated with histological type, lymphatic invasion, lymph node metastasis, and microvessel density. No association was identified between VEGF-D immunoreactivity and clinicopathological variables. These results suggest that VEGF-C is a dominant regulator of lymphangiogenesis in early-stage human gastric carcinoma.     

VEGF-C和VEGF-D在胰腺癌淋巴转移中的意义

目的 分析胰腺癌肿瘤中心和肿瘤周边组织中血管内皮生长因子(VEGF)-C、VEGF-D与微血管密度(MVD)、微淋巴管密度(MLVD)的关系,探讨VEGF-C、VEGF-D在胰腺癌淋巴结转移及发展中的意义.方法 免疫组织化学法检测30例胰腺癌组织中VEGF-C、VEGF-D、VEGF受体(VEGFR)-3、CDM蛋白的表达情况.结果 30例胰腺癌中肿瘤周边部位VEGF-C、VEGF-D蛋白阳性率分别为73.3%和56.7%,显著高于肿瘤中心部位(30.0%和16.7%,P<0.01).VEGF-C、VEGF-D高表达的肿瘤周边部位淋巴结转移、淋巴管和血管浸润显著增加(P<0.01).VEGF-C蛋白阳性组MVD高于阴性组,MLVD显著高于阴性组(P<0.01),淋巴结转移增多;VEGF-D蛋白阳性组与阴性组相比MVD无变化(P=0.07),MLVD高于阴性组(P<0.01),淋巴结转移增加.结论 胰腺癌中肿瘤周边区域中VEGF-C、VEGF-D的表达与患者淋巴结转移显著相关,并介导其淋巴管生成;而VEGF-C可能主要参与胰腺癌的血管生成和淋巴管生成的调节,VEGF-D可能仅参与其淋巴管生成的调节.     

血管内皮生长因子-D在胃癌中表达的意义

Objective： To investigate the expression of vascular endothelial growth factor D （VEGF-D） in gastric cancer and its relationship with lymph node metastasis. Methods： 100 cases of gastric carcinoma tissues （50 cases with lymph node metastasis, 50 cases negative） and 30 cases of normal gastric tissues were gathered to detect the expressions of VEGF-C and D proteins by immunohistochemistry. Results： VEGF-C and D were revealed in cytoplasm of gastric carcinoma tissues and normal gastric tissues. The positive rates of VEGF-C and D expressions were significantly higher in gastric cancer tissues than those in the normal ones （51%, 60% vs 10%, 20% respectively; both P 〈 0.05）. There were significant correlations between the positive expression of VEGF-D and lymph node metastasis, lymphatic invasion, positive expression of VEG F-C, but not with tumour size, tissue differentiation, and venous invasion. Conclusion： The expression of VEGF-D is closely related to lymph node metastasis in gastric carcinoma.     

Impact of vascular endothelial growth factor-C and -D expression in human pancreatic cancer: its relationship to lymph node metastasis.

PURPOSE: The aim of this study was to evaluate the expression of vascular endothelial growth factor (VEGF)-C and -D in pancreatic cancer and to reveal its relation to lymph node metastasis. EXPERIMENTAL DESIGN: Formalin-fixed, paraffin-embedded blocks were obtained from 58 patients with pancreatic head cancer. All of the patients underwent a curative resection. The total number of resected lymph nodes was 1,058. The expressions of VEGF-C and -D were evaluated by immunohistochemical staining. To evaluate the relation to lymph node metastasis, the expressions of VEGF-C and -D between the marginal and central portions in the tumor were compared. When >25% of the tumor cells showed distinct staining, the portion was judged as high expression. RESULTS: The two groups with high expression of VEGF-C (P = 0.015) and VEGF-D (P = 0.020) in the marginal portion had a significantly higher incidence of lymph node metastasis compared with the groups with low expression, respectively. Furthermore, the group with high expression of both VEGF-C and -D in the marginal portion had a higher incidence of lymph node metastasis compared with the group with low expression (P = 0.007). The 5-year survival rate of patients with high expression of both VEGF-C and -D in the marginal portion was significantly lower than that of patients with low expression of both VEGF-C and -D (P = 0.017). CONCLUSIONS: VEGF-C and -D expression in tumor cells in the marginal portion of the tumor significantly associated with lymphatic metastasis and prognosis in patients with pancreatic head cancer.     

Tumor-induced activation of lymphatic endothelial cells via vascular endothelial growth factor receptor-2 is critical for prostate cancer lymphatic metastasis

Prostate cancer disseminates initially and primarily to regional lymph nodes. However, the nature of interactions between tumor cells and lymphatic endothelial cells (LEC) is poorly understood. In the current study, we have isolated prostate LECs and developed a series of two-dimensional and three-dimensional in vitro coculture systems and in vivo orthotopic prostate cancer models to investigate the interactions of prostate cancer cells with prostate LECs. In vitro, highly lymph node metastatic prostate cancer cell lines (PC-3 and LNCaP) and their conditioned medium enhanced prostate LEC tube formation and migration, whereas poorly lymph node metastatic prostate cancer cells (DU145) or normal prostate epithelial cells (RWPE-1) or their conditioned medium had no effect. In vivo, the occurrence of lymphatic invasion and lymph node metastasis was observed in PC-3 and LNCaP xenografts but not in DU145 xenografts. Furthermore, vascular endothelial growth factor (VEGF) receptor (VEGFR)-2 is expressed by prostate LECs, and its ligands VEGF-A, VEGF-C, and VEGF-D are up-regulated in highly lymph node metastatic prostate cancer cells. Recombinant VEGF-A and VEGF-C, but not VEGF-C156S, potently promoted prostate LEC tube formation, migration, and proliferation in vitro, indicating that signaling via VEGFR-2 rather than VEGFR-3 is involved in these responses. Consistent with this, blockade of VEGFR-2 significantly reduced tumor-induced activation of LECs. These results show that the interaction of prostate tumor cells with LECs via VEGFR-2 modulates LEC behavior and is related to the ability of tumor cells to form lymph node metastases.     

Role of vascular endothelial growth factor receptor-3/Flt-4 in early-stage cervical cancer

The present study was designed to investigate the role of vascular endothelial growth factor receptor (VEGFR)-3/Flt-4 in the early stages of cervical cancer. VEGFR-3/Flt-4 expression, vascular endothelial growth factor (VEGF)-C and VEGF-D in the early stages (Ia-IIa) of cervical cancer in 41 patients was examined by immunohistochemical analysis. Additionally, the VEGFR-3/Flt-4-marked vascular density (MVD) was examined and the relationship of these factors with clinicopathological factors was analyzed. VEGFR-3/Flt-4 was found to be expressed in lymphatic endothelial cells and, to a certain extent, in vascular endothelial cells. The VEGFR-3/Flt-4-positive vessels were largely distributed in the stroma surrounding the tumor tissues, and these vessels were morphologically divided into blood and lymphatic vessels, respectively. Cancer cells were found to express VEGF-C, VEGF-D and VEGFR-3/Flt-4, and their positive expression rate was 48.7% (20/41), 58.5% (24/41) and 63.4% (26/41), respectively. VEGFR-3/Flt-4 expression in the cancer cells of the cervical cancer patients in our study was found to be correlated to the clinical stage, lymph node metastasis, lymphatic invasion and expression of VEGF-C and VEGF-D. However, this expression was unrelated to menstrual status, histological grade and histological classification. MVD was correlated to the clinical stage and expression of VEGF-C and VEGF-D, but was unrelated to menstrual status, histological grade, histological classification, lymph node metastasis and lymphatic invasion. In conclusion, VEGFR-3/Flt-4 plays an important role in the early stages of cervical cancer.     

The expression of vascular endothelial growth factor C and its receptors in non-small cell lung cancer.

Expression of vascular endothelial growth factor (VEGF)-C and that of its receptors were assessed in non-small cell lung cancer. Immunohistochemistry revealed positive VEGF-C expression in 38.7% (24/62) of the patients studied. A significant positive correlation was found between VEGF-C in cancer cells and VEGF receptor-3 (VEGFR-3) in vascular endothelial cells, but not between VEGF-C in cancer cells and VEGFR-2 in endothelial cells. In this cohort of lung cancer patients, VEGF-C expression was significantly associated with lymph node metastasis, lymphatic vessel invasion, and worse outcomes after the operation. Although the independent prognostic impact of VEGF-C and VEGFR-3 was not clear, VEGFR-2 expression in endothelial cells retained the independency as the prognostic indicator. In light of these findings, we conclude that VEGF-C plays an important role in lymphatic invasion/metastasis and tumour progression in non-small cell lung cancer.     

血管内皮生长因子D在胃癌中表达的意义

 目的 研究血管内皮生长因子D（VEGF-D）在胃癌组织内表达的意义。方法 选择手术后的100例胃癌（区域淋巴结转移阳性和阴性病例各50例），30例正常胃黏膜设为对照组。利用免疫组化EnVision法，检测研究对象胃组织内VEGF-D和VEGF-C的表达。分析VEGF-D阳性表达与临床病理参数和VEGF-C阳性表达的关系。结果 VEGF-C，VEGF-D的阳性染色为棕黄色，主要位于肿瘤细胞的胞质，局灶性或弥散性分布。VEGF-C，VEGF-D阳性表达以（+）或（++）为主；胃癌VEGF-C、VEGF-D阳性表达率分别为51 %和60 %；对照组分别为10 %和20 %（P＜0.05）；VEGF-D阳性高表达（++）与区域淋巴结转移、淋巴管受侵和VEGF-C阳性表达相关，与肿瘤最大径、肿瘤位置、肿瘤浸润深度、肿瘤肉眼分型、肿瘤组织分化、肿瘤血管受侵无关。结论 VEGF-D在胃癌淋巴道转移过程中具有较重要的作用。     

VEGF121 promotes lymphangiogenesis in the sentinel lymph nodes of non-small cell lung carcinoma patients

BACKGROUND: The sentinel lymph node (SLN) concept is that lymphatic flux from a primary tumor initially flows into a SLN. The mechanism mediating tumor metastasis within SLNs remains largely unknown; however, primary tumors overexpressing vascular endothelial growth factor (VEGF)-A appear to induce SLN lymphangiogenesis prior to metastasis in animal model. Our aim was to further investigate the capacity of VEGFs to induce lymphangiogenesis within SLNs and to assess their role in SLN metastasis in non-small cell lung carcinoma (NSCLC). METHODS: Real-time quantitative RT-PCR was used to assess expression of mRNAs encoding several VEGFs (VEGF121, VEGF165, VEGFR1, VEGFR2, VEGFR3, VEGF-C and VEGF-D) in resected lymph node specimens from 35 NSCLC patients, after which we compared their expression SLNs and non-SLNs. In addition, expression of the lymphatic endothelium-specific hyaluronan receptor (LYVE)-1 was used to assess lymphangiogenesis in SLNs and non-SLNs. RESULTS: Immunohistochemical staining revealed substantial expression of LYVE-1 in SLNs. Moreover, levels LYVE-1 mRNA were significantly higher in SLNs than non-SLNs (P<0.05), as were levels of VEGF121 and VEGFR2 mRNA (P<0.01 and P=0.02, respectively). In addition metastasis-positive SLNs showed significantly higher levels of VEGF121, VEGF-C and VEGF-D mRNA than metastasis-negative SLNs (P<0.001, P=0.01 and P=0.01, respectively), and VEGF121 induced the proliferation of lymphatic endothelial cells (P<0.01). CONCLUSIONS: Our findings suggest that active lymphangiogenesis is ongoing within SLNs from NSCLC patients, even before metastasis. This lymphangiogenesis may be promoted by upregulation of VEGF121, which may in turn act in part via induction of VEGF-C.     

血管内皮生长因子C、D在鼻咽癌组织中的表达及其临床意义

背景与目的:血管内皮生长因子C(vascular endothelial growth factor-C,VEGF-C)和D(vascular endothelial growth factor-D,VEGF-D)是目前已鉴定出的淋巴管生长因子,有研究表明肿瘤组织中VECF-C或VEGF-D过表达与淋巴转移有关.本研究旨在探讨鼻咽癌组织中VEGF-C和VEGF-D的表达情况及其临床意义.方法:采用免疫组化SP法检测66例鼻咽癌组织中VEGF-C和VEGF-D的表达情况,同时检测血管内皮生长因子受体3(vascular endotheliaI growth factor receptor-3,VEGFR-3)和CD34染色情况并计数微淋巴管密度(lymphatic microvessel density,LMVD)和微血管密度(microvessel density,MVD).结果:VEGF-C高表达率在鼻咽癌组织中(54.5%)较鼻咽非肿瘤组织中(26.3%)高(P＜0.05);鼻咽癌伴区域淋巴结转移或T分期晚者,VEGF-C高表达率增高,单因素及多因素Logistic回归分析均表明区域淋巴结转移与VEGF-C高表达相关(P＜0.05),但VEGF-C高表达与性别、年龄、5年生存率、LMVD、MVD等因素无关(P＞0.05).VEGF-D阳性表达率在鼻咽癌组织中(69.7%)较鼻咽非肿瘤组织中(42.1%)高(P＜0.05);鼻咽癌中VEGF-D阳性表达与性别、年龄、T分期、区域淋巴结转移、LMVD、MVD等因素无关(P＞0.05),但与VEGF-C高表达显著正相关(P＜0.01),VEGF-D阳性表达者5年生存率(50.0%)显著低于VEGF-D不表达者(85.0%)(P＜0.01).结论:鼻咽癌中VEGF-C高表达与区域淋巴结转移密切相关;VEGF-D阳性表达与区域淋巴结转移无关,但与VEGF-C高表达正相关,且与5年生存率密切相关.     

Pathways targeting tumor lymphangiogenesis.

Tumor metastasis to sentinel lymph nodes represents the first step of tumor dissemination in most human cancers and serves as a major prognostic indicator for disease progression. Recent studies have revealed that tumors can actively induce the formation of lymphatic vessels, and that tumor lymphangiogenesis is correlated with lymph node metastasis in experimental cancer models and in several types of human cancers. Metastatic tumor cells may continue to promote lymphatic vessel growth even after their metastasis to sentinel lymph nodes, likely promoting further cancer spread. Vascular endothelial growth factor-C (VEGF-C) and VEGF-D were the first specific lymphangiogenesis factors identified, acting predominantly via VEGF receptor-3 (VEGFR-3) that is expressed by lymphatic endothelial cells, and a large number of clinical studies have shown a correlation between tumor expression of VEGF-C or VEGF-D and lymph node metastasis. VEGFR-3 activation promotes lymphatic endothelial cell proliferation, migration, and survival via the extracellular signal-regulated kinase 1/2, the phosphatidylinositol 3-kinase/AKT, and the c-Jun NH(2)-terminal kinase 1/2 pathways. Additional tumor lymphangiogenesis factors have been recently identified, including VEGF-A. Importantly, blockade of the VEGFR-3 pathway by specific antibodies, by soluble receptor constructs, and by small molecule kinase inhibitors efficiently inhibits experimental tumor lymphangiogenesis and metastasis and might also represent a novel therapeutic avenue for the treatment of human cancers.     

Serum vascular endothelial growth factor-C level in patients with primary nonsmall cell lung carcinoma: a possible diagnostic tool for lymph node metastasis

BACKGROUND: The authors measured circulating vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor (VEGF) levels in patients with primary nonsmall cell lung carcinoma and assessed its usefulness as a diagnostic tool for determining lymph node metastasis. METHODS: Ninety-two patients with nonsmall cell lung carcinoma and 58 patients with benign tumors of the lung were included in the current study, as well as 42 healthy control patients. Circulating VEGF-C and VEGF levels were assessed by enzyme-linked immunosorbent assay. RESULTS: Serum VEGF-C concentration was higher in patients with lung carcinoma than in patients with benign tumors or in healthy control patients. Patients with lymph node metastasis revealed higher serum VEGF-C and VEGF concentrations than those without. The median level of VEGF-C and VEGF according to lymphatic vessel invasion and venous invasion was higher in the group with invasion than in the group without. These differences were most significant among patients with VEGF-C with lymphatic vessel invasion (P = 0.0066 vs. P = 0.026) and in patients with VEGF with venous invasion (P = 0.19 vs. P = 0.011). Serum VEGF-C levels reached a sensitivity of 79% and a specificity of 72% with a cutoff value of 1756.0 pg/mL, whereas VEGF levels reached 68% sensitivity and 70% specificity at 327.8 pg/mL. If 92 patients were divided into 4 groups according to the combination of VEGF-C and VEGF levels, the positive predictive value was 84.2%, the negative predictive value was 95.8%, and accuracy was 93.1%. CONCLUSIONS: Circulating VEGF-C levels may provide additional information for distinguishing between the absence and presence of lymph node metastasis in patients with lung carcinoma.