Expression of vascular endothelial growth factor-C and angiogenesis in esophageal squamous cell carcinoma

AIM: To investigate the expression of vascular endothelial growth factor-c (VEGF-C) mRNA and microvessel density (MVD) in human esophageal squamous cell carcinoma (ESCC) and its relationship with clinical significance. METHODS: Specimens obtained from 43 patients undergoing surgical resection for ESCC were used in this study. The expression of VEGF-C mRNA was examined by in situ hybridization. Tumor MVD was determined immunohistochemically with anti-CD31 antibody and estimated by image analysis. Ten sections of adjacent normal mucosa were also examined. RESULTS: VEGF-C mRNA expression was detected in cytoplasm of carcinoma cells. Of the 43 ESCC patients studied, 18 cases (41.9%) were positive for VEGF-C mRNA. No VEGF-C mRNA expression was observed in normal esophageal mucosa. VEGF-C mRNA expression correlated significantly with lymph node metastasis, TNM stage and depth of invasion (P < 0.05). Furthermore, histological grade (differentiation) tended to correlate with VEGF-C mRNA expression, but was not statistically significant (P > 0.05). In tumor lesions, the MVD was significantly greater than that in normal esophageal mucosa. MVD correlated significantly with lymph node metastasis, TNM stage and depth of invasion (P < 0.05), but not with histological grade (differentiation) (P > 0.05). Lesions with VEGF-C mRNA expression had a significantly higher MVD than that of those without VEGF-C mRNA expression (P < 0.05). CONCLUSION: VEGF-C plays a role in lymphatic metastasis via lymphangiogenesis and angiogenesis in ESCC. VEGF-C is one of the important predictors of the biological behavior in ESCC.     

COX-2 expression is correlated with VEGF-C, lymphangiogenesis and lymph node metastasis in human cervical cancer

Lymphangiogenesis has been shown to promote lymph node metastasis in cancers, making it an important target in cancer therapy. Vascular endothelial growth factor (VEGF)-C is upregulated in various tumors/cancers and is one of the most potent growth factors for inducing lymphangiogenesis and promoting lymph node metastasis (LNM). Likewise, cyclooxygenase (COX)-2 plays major roles in carcinogenesis, tumor growth and metastasis via multiple mechanisms including inactivation of host antitumor immunity and promotion of tumor cell migration, tumor cell invasiveness and tumor-associated angiogenesis and lymphangiogenesis. We previously demonstrated an association between COX-2 and VEGF-C in an in vitro model of lung cancer. However, little is known about the regulation of VEGF-C by COX-2 in cervical cancer. In this study, we measured the COX-2 and VEGF-C expressions by immunohistochemistry in 23 LNM-positive and 20 LNM-negative cervical cancer specimens. We then examined the correlations among the expressions and the lymphatic microvessel density (LMVD) and ultrastructural changes to the lymphatic vessel walls by enzyme histochemical staining and electron microscopy. In addition, we used the HeLa cervical cancer cell line to explore the in vitro regulation of VEGF-C by COX-2 and its metabolite, PGE₂, using siRNA-mediated gene silencing and EP receptor blockade. The LNM-positive specimens exhibited significantly higher VEGF-C expression, COX-2 expression and LMVD than the LNM-negative specimens. Furthermore, there were strong correlations between the levels of COX-2 expression and the levels of VEGF-C expression and secretion and a significant positive association between the LMVD and LNM. siRNA-mediated knockdown of COX-2 expression inhibited VEGF-C mRNA expression while EP1 and EP4 receptor antagonists reduced the VEGF-C protein level and tyrosine phosphorylation of Src kinase. Moreover, inhibition of Src kinase with the tyrosine kinase inhibitor PP1 attenuated VEGF-C expression. Collectively, our data provide evidence for a clinical association between COX-2 and VEGF-C expressions in cervical cancer. EP1 and EP4 receptors may be involved in the COX-2-mediated regulation of VEGF-C protein and mRNA expressions. Src may be a downstream mediator of EP1 and EP4 receptors. COX-2 inhibition may diminish LNM by suppressing VEGF-C-mediated lymphangiogenesis.     

Vascular endothelial growth factor-C (VEGF-C) is a more specific risk factor for lymph node metastasis than VEGF-D in submucosal colorectal cancer

BACKGROUND/AIMS: It is useful to decide whether lymphatic involvement or lymph node metastasis exists before polypectomy or operation in submucosal colorectal cancer. Whether vascular endothelial growth factor-C (VEGF-C) or VEGF-D could predict lymph node metastasis and lymphatic involvement is uncertain. METHODOLOGY: Expression of the VEGF-C and VEGF-D in human submucosal colorectal cancers was investigated in paraffin-embedded stepwise sections by means of immunohistochemistry, and the correlation between immunohistochemical expression pattern and clinicopathological features was also evaluated. RESULTS: The results showed that VEGF-C overexpression correlated with lymphatic involvement (P = 0.01) and lymph node metastasis (P = 0.02), but VEGF-D overexpression did not correlate significantly. In multivariate analysis lymphatic invasion was the predictive factor (P = 0.0129), but VEGF-C positivity was not predictive (P = 0.3437). CONCLUSIONS: These results may suggest that VEGF-C is a more specific risk factor for lymph node metastasis than VEGF-D in submucosal colorectal cancer.     

鼻咽癌组织中COX-2和VEGF-C的表达及意义

目的探讨鼻咽癌组织中环氧合酶（COX）-2及血管内皮生长因子（VEGF）-C的表达变化及意义。方法采用免疫组化SP法检测58例鼻咽癌组织和10例正常鼻咽黏膜组织中COX-2及VEGF—C的表达，分析其与淋巴结转移的关系。结果鼻咽癌组织中COX-2、VEGF-C的阳性表达率分别为65．5％（38／58）、77．5％（45／58），显著高于正常鼻咽黏膜组织的10％（1／10）、10％（1／10），P均〈0．05；COX-2、VEGF—C的表达与鼻咽癌的淋巴结转移及分化程度相关，P均〈0．05；鼻咽癌中COX-2、VEGF—C的表达呈正相关，r=0．491，P〈0．01．结论鼻咽痛中COX-2、VEGF—C表试均增高，二者共同促进鼻咽癌的淋巴结转移。     

microRNA-101和环氧合酶-2在胃癌中的表达及其临床意义

背景：microRNAs是一类对靶基因表达具有转录后调控作用的非编码小RNA。microRNA-101（miR-101）在多种恶性肿瘤中呈低表达,而过表达外源性miR-101可发挥肿瘤抑制作用。前期体内、外实验发现外源性miR-101可抑制胃癌细胞增殖和环氧合酶-2（COX-2）表达。目的：检测胃癌组织中的miR-101、COX-2表达并探讨其临床意义。方法：收集手术切除胃癌组织和配对癌旁非癌组织标本30例,以实时荧光定量RT—PCR检测miR-101、COX-2mRNA表达,分析两者间以及两者与胃癌主要临床病理特征的关系。结果：绝大多数胃癌组织中miR-101表达低下,COX-2mRNA则呈过表达。胃癌组织与癌旁非癌组织间miR-101、COX-2mRNA表达量差异显著（P〈0．01）,且两者表达在癌组织和癌旁组织中均呈负相关（癌组织：r=-0．767,P=0．000;癌旁组织：r=-0．718,P=0．000）。TNMⅢ、IV期胃癌和伴淋巴结转移的胃癌中,miR-101表达分别显著低于TNMI、Ⅱ期病例和无淋巴结转移病例（P〈0．05）,COX-2mRNA表达分别显著高于TNMI、Ⅱ期病例和无淋巴结转移病例（P〈0．05）。结论：miR-101与COX-2之间的负相关性可能有助于胃癌的临床诊断;miR-101表达低下伴COX-2过表达与胃癌临床进展和转移有关,对预后判断有一定参考价值。     

OPN和VEGF-C在胃癌中的表达及其临床意义

目的:评价骨桥蛋白(osteopontin,OPN)和血管内皮生长因子C(vascular endothelial growth factor C,VEGF-C)在胃癌中表达及与临床病理参数之间相关性,进一步探讨二者的共表达在胃癌淋巴结转移中的机制.方法:收集复旦大学附属中山医院手术切除胃癌组织标本及患者相关临床资料,对纳入研究的93例胃癌原发灶标本采用免疫组织化学染色法(EnVision二步法),检测OPN与VEGF-C蛋白的表达.结果:93例胃癌组织中OPN与VEGF-C的阳性表达率分别为64.5%(60/93)和69.9%(65/93),在非肿瘤性胃黏膜中均未见其阳性表达;OPN和VEGF-C的表达与胃癌浆膜侵犯、TNM分期以及淋巴结转移呈明显相关(P<0.05).蛋白之间的相关性分析显示,OPN与VEGF-C之间存在正相关(r=0.493,P<0.01).结论:联合检测OPN与VEGF-C有助于阐述胃癌发生发展、浸润转移的机制,OPN可能通过上调VEGF-C的表达促进胃癌的淋巴结转移.     

Involvement of cyclooxygenase-2 and vascular endothelial growth factor in vascularization and lymph node metastasis of colorectal cancers with submucosal invasion

BACKGROUND AND AIMS: Although patients with early colorectal cancer invading the submucosa (CRC-sm) may be treated with endoscopic mucosal resection alone, they generally undergo additional surgery because of the risk of lymph node metastasis. The aims of the present study were to examine the roles of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) in tumor vascularization and to investigate whether COX-2 and VEGF expression and tumor vascularity are useful markers for predicting lymph node metastasis in CRC-sm. METHODS: Twenty-seven resected specimens of CRC-sm with lymph node dissection were examined, and expression of COX-2 and VEGF was evaluated immunohistochemically and scored. Microvessel density (MVD) in CRC-sm tissues was estimated using a Macscope system after CD34 immunostaining. The relationships among clinicopathological parameters, COX-2 and VEGF expression, and MVD in CRC-sm tissues were then analyzed. RESULTS: Scores for COX-2, VEGF and MVD were all significantly higher in patients with CRC-sm with lymphatic invasion or lymph node metastasis. COX-2 score (P < 0.0001) and VEGF score (P = 0.035) were significantly correlated with MVD in CRC-sm tissues. In addition, COX-2 score was significantly correlated with VEGF score in the CRC-sm specimens examined. CONCLUSIONS: Both COX-2 and VEGF are involved in tumor vascularization in CRC-sm. COX-2 expression, VEGF expression, and MVD are possible markers for predicting lymph node metastasis in patients with CRC-sm, and use of COX-2 expression may be clinically practical.     

Vascular endothelial growth factor C and D expression correlates with lymph node metastasis and poor prognosis in patients with resected esophageal cancer

Vascular endothelial growth factors C (VEGF-C) and D (VEGF-D) are important lymphangiogenic factors in human cancers. We studied the expression of VEGF-C and VEGF-D using immunohistochemistry in 73 resected esophageal cancer specimens, and correlated the results with patient clinicopathologic features and survival. High expression of VEGF-C was identified in 40 (54.7%) patients, and it correlated positively with histological grade (p=0.038), tumor stage (p=0.01), depth of tumor invasion (p=0.036) and lymph node metastasis (p=0.001). In 48 of 73 (65.7%) tumors, the VEGF-D protein was also expressed at high levels. VEGF-D immunoreactivity significantly correlated with tumor location (p=0.027), size of tumor (p=0.015), histological grade (p=0.02), depth of invasion (p=0.001) and lymph node metastasis (p=0.018). In logistic multivariate analysis, high expression of VEGF-C (OR 1.941, 95% CI 1.263-7.289, p=0.024) was associated with lymph node metastasis. Calculating the prognostic relevance revealed that both VEGF-C and VEGF-D correlated with decreased overall survival (p=0.01, p=0.003), disease free survival (p=0.02, p=0.006), and cancer-specific survival (p=0.03, p=0.005). In conclusion, our results suggest that high levels of both VEGF-C and VEGF-D proteins are associated with lymph node involvement, and that VEGF-C expression is an independent predictor of risk for lymph node metastasis in esophageal cancer. In locally advanced disease, overexpression of VEGF-C and VEGF-D may be useful in identifying patients who are more likely to have a poor prognosis even after curative resection.     

Expression of vascular endothelial growth factor-C in benign and malignant thyroid tumors

In contrast to vascular endothelial growth factor (VEGF), which stimulates angiogenesis, VEGF-C is thought to stimulate lymphangiogenesis. The role of VEGF-C in thyroid cancer pathogenesis has not been clarified. One might expect a different pattern of VEGF-C expression in the various types of thyroid cancer because of their different means of metastases. In this investigation, we determined whether the differential expression of VEGF-C might explain the different propensity to lymph node metastasis in thyroid cancers. One hundred eleven normal and neoplastic thyroid tissues were analyzed by real-time quantitative PCR. Papillary thyroid cancers had a higher VEGF-C expression than other thyroid malignancies (P < 0.0005 ANOVA). Among the normal thyroid tissues from patients with malignant or benign thyroid diseases, there was no significant difference in VEGF-C expression. Paired comparison of VEGF-C expression between thyroid cancers and normal thyroid tissues from the same patients showed a significant increase of VEGF-C expression in papillary thyroid cancer (1.10 +/- 0.41 vs. 0.70 +/- 0.13; P = 0.001) and a significant decrease of VEGF-C expression in medullary thyroid cancer (0.11 +/- 0.13 vs. 0.78 +/- 0.29; P = 0.001). In contrast, there was no significant difference of VEGF-C expression between cancer and normal tissues in other types of thyroid cancer. In summary, VEGF-C expression is increased in papillary thyroid cancer, compared with paired normal thyroid tissues, but not in other thyroid cancers that are also prone to lymph node metastasis. The lymphangiogenic role of VEGF-C in thyroid cancers therefore appears to be complex and other factors are likely to be also involved.     

喉癌组织中VEGF-C及其受体VEGFR-3和D2-40的表达及意义

目的 观察血管内皮生长因子C（VEGF-C）及其受体（VEGFR-3）和D2-40在喉癌组织中的表达情况,探讨其在喉癌淋巴道转移过程中的作用及可能机制.方法 选择喉癌患者的癌组织、癌旁正常组织标本各40份,同期选择正常喉组织标本19份作对照.采用免疫组织化学法检测各标本中VEGF-C、VEGFR-3和D240的表达.结果 VEGF-C、VEGFR-3及D2-40在喉癌组织、癌旁组织及正常喉组织中均有表达,且其表达强度呈下降趋势（H分别为38.400、32.502,P均＜0.01）;在喉癌组织中,VEGF-C、VEGFR-3及D2-40的表达与淋巴转移、临床分期及临床分型有关（P均＜0.05）,与病理分级、肿瘤大小、吸烟量、年龄和性别无关（P均＞0.05）.喉癌组织中VEGF-C与VEGFRd的表达呈正相关（r=0.882,P＜0.01）.结论 VEGF-C、VEGFR-3及D2-40在喉癌组织中的表达均明显高于癌旁组织及正常喉组织,其高表达可能与喉癌的发生、发展有关.     

Cancer stem cell marker ALDH1 expression is associated with lymph node metastasis and poor survival in esophageal squamous cell carcinoma: a study from high incidence area of northern China

Tumor recurrence and metastasis is the leading cause of death in esophageal squamous cell carcinoma (ESCC). Cancer stem cell (CSC) may be responsible for tumor growth and maintenance of aggressive behavior. Aldehyde dehydrogenase 1 (ALDH1) has been proposed as one of the possible candidates for a CSC marker. The expression of ALDH1 may be correlated with the clinicopathologic factor and clinical outcome of patients with ESCC. The purpose of this study was to investigate the expression of ALDH1 protein in human ESCC tissues, and evaluated the clinical implication of ALDH1 expression for these patients. All 79 patients who underwent esophagectomy for ESCC between January 2005 and June 2006 were enrolled in this study. The expression of ALDH1 in ESCC and adjacent noncancerous tissues was analyzed by immunohistochemistry. ALDH1 was mainly expressed in ESCC cell nucleus. For the 79 ESCC patients, increased nuclear accumulation of ALDH1 was found in 12 (15.2%) specimens. ALDH1 expression was correlated with poor histological differentiation (P= 0.003), lymph node metastasis (P= 0.011), and late pathologic TNM classification (pTNM) staging (P= 0.003). Patients in ALDH1 positive group had a significantly poor 5-year overall survival than those in the negative group (8.3% vs. 52.2%, P= 0.025). We have demonstrated for the first time that the CSC marker, ALDH1, is expressed in human ESCC. The expression of ALDH1 protein in nucleus of the ESCC is significantly associated with lymph node metastasis and poor survival. Our results highly indicate the involvement of ALDH1 in the aggressive behavior of ESCC.     

Expression of vascular endothelial growth factor C and chemokine receptor CCR7 in gastric carcinoma and their values in predicting lymph node metastasis

AIM: To study the expression of vascular endothelial growth factor C (VEGF-C) and chemokine receptor CCR7 in gastric carcinoma and to investigate their associations with lymph node metastasis of gastric carcinoma and their values in predicting lymph node metastasis. METHODS: The expression of VEGF-C and CCR7 in gastric carcinoma tissues obtained from 118 patients who underwent curative gastrectomy was examined by immunohistochemistry. Among these patients, 39 patients underwent multi-slice spiral CT (MSCT) examination. RESULTS: VEGF-C and CCR7 were positively expressed in 52.5 and 53.4% of patients. VEGF-C expression was more frequently found in tumors with lymph node metastasis than those without it (P<0.001). VEGF-C expression was also closely related to lymphatic invasion (P<0.001), vascular invasion (P<0.01), and TNM stage (P<0.001). However, there was no significant correlation between VEGF-C expression and age at surgery, gender, tumor size, tumor location, Lauren classification, and depth of invasion. CCR7 expression was significantly higher in patients with lymph node metastasis compared with those without lymph node metastasis (P<0.001) and was also associated with tumor size (P<0.01), depth of invasion (P<0.001), lymphatic invasion (P<0.001), and TNM stage (P<0.001). However, the presence of CCR7 had no correlation to age at surgery, gender, tumor location, Lauren classification, and vascular invasion. Among the 39 patients who underwent MSCT examination, only CCR7 expression was related to lymph node metastasis determined by MSCT (P<0.05). In the current retrospective study, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of VEGF-C and CCR7 expression in the diagnosis of lymph node metastasis for patients with gastric carcinoma were 73.8%, 70.2%, 72.6%, 71.4% and 72.0%, and 82.0%, 77.2%, 79.4%, 80.0% and 79.7%, respectively. After subdivision according to the combination of VEGF-C and CCR7 expression, receiver operating characteristic (ROC) analysis showed that the accuracy of the combined examination of VEGF-C and CCR7 expression in predicting lymph node metastasis was relatively high (area under ROC curve [Az]=0.83). CONCLUSION: The expression of VEGF-C and CCR7 is related to lymph node metastasis of gastric carcinoma and both of them may become new targets for the treatment of gastric carcinoma. Furthermore, the combined examination of VEGF-C and CCR7 expression in endoscopic biopsy specimens may be useful in predicting lymph node metastasis of gastric carcinoma and deciding the extent of surgical lymph node resection.     

VEGFR-3信号途径相关蛋白在胃癌中的表达及意义

目的研究血管内皮生长因子受体（VEGFR）-3信号途径相关蛋白C、D（VEGF—C、VEGF—D）在胃癌中的表达并探讨其与淋巴结转移的关系。方法80例胃癌手术病例分为淋巴结阳性组（n=48）和淋巴结阴性组（n=32）,另设胃溃疡病例为对照组（n=10）。采用ELISA技术进行血清VEGF—C和VEGF-D水平检测,然后应用免疫组织化学EnVisionTM两步法检测胃溃疡组织和胃癌标本VEGF—C、VEGF-D表达。结果胃癌患者血清VEGF-C和VEGF—D水平明显高于对照组（P〈0．05）,胃癌患者血清VEGF-C水平与淋巴结转移有关（P〈0．05）;胃癌组织中VEGF—C、VEGF—D阳性表达率均高于对照组（P〈0．05）,伴淋巴结转移的胃癌组织VEGF-C阳性表达率较无淋巴结转移者更高（P〈0．01）。结论血清VEGF—C可作为胃癌的标记物,对术前判定淋巴结转移具有一定的临床价值。     

Impact of lymph node micrometastasis in hilar bile duct carcinoma patients

AIM: To immunohistochemically examine micrometastasis and VEGF-C expression in hilar bile duct carcinoma (HBDC) and to evaluate the clinical significance of the results. METHODS: A total of 361 regional lymph nodes from 25 patients with node-negative HBDC were immunostained with an antibody against cytokeratins 8 and 18 (CAM 5.2), and immunohistochemical staining of VEGF-C was performed in 34 primary resected tumors. RESULTS: Lymph node micrometastasis was detected in 6 (24%) of the 25 patients and 10 (2.8%) of the 361 lymph nodes. Patients with micrometastasis showed significantly poorer survival rates than those without (P=0.025). VEGF-C expression was positive in 17 (50%) of 34 HBDC, and significantly correlated with lymph node metastasis (P=0.042) and microscopic venous invasion (P=0.035). CONCLUSIONS: It is suggested that immunohistochemically detected lymph node micrometastasis has an impact on the outcome of HBDC. VEGF-C expression is highly correlated with lymph node metastasis in HBDC and might therefore be a useful predictor.     

Persistent CXCR4 expression after preoperative chemoradiotherapy predicts early recurrence and poor prognosis in esophageal cancer

INTRODUCTIONThe migration of tumor cells to a secondary site from their primary location is a crucial issue in cancer metastasis. Recently, a novel “homing” signaling mechanism has been proposed, in which target organs produce and release specific chemo…     

环氧合酶-2和血管内皮生长因子-C与胃癌淋巴管转移

目的　研究环氧合酶 2 (COX 2 )和血管内皮生长因子 (VEGF) C在胃癌组织中的表达及相关性 ,探讨二者在胃癌淋巴管生成和转移中的作用。方法　采用免疫组化方法和逆转录 PCR技术 ,分别检测了 6 4例胃癌石蜡组织中COX 2和VEGF C的表达及其中 2 2例胃癌新鲜组织中二者mRNA的表达。结果　 2 2例胃癌新鲜组织中COX 2和VEGF CmRNA表达阳性率分别为 82 %和73% ,其表达均高于相应的癌旁正常组织 (P <0 0 0 1) ,与 6 4例胃癌石蜡标本COX 2和VEGF C的表达结果较一致。且COX 2和VEGF C表达之间存在明显关联性 (P <0 0 5 )。二者在癌组织中的高表达与肿瘤浸润深度、淋巴结转移等密切相关 (P <0 0 5 )。结论　胃癌组织中有COX 2和VEGF C的高表达 ,而COX 2可能参与VEGF C淋巴管生成通路 ,它们的表达可能在胃癌淋巴管浸润和转移中发挥重要作用     

Association between Bmi1 and clinicopathological status of esophageal squamous cell carcinoma

AIM： To investigate the clinicopathological roles of Bmil in esophageal squamous cell carcinoma （ESCC）.METHODS： Quantitative real-time polymerase chain reaction and immunohistochemical staining for Broil were performed in cancerous and adjacent non-cancerous paraffin-embedded esophageal specimens.RESULTS： The Bmil expression level was unaffected by gender and age. The level of Broil mRNA in ESCC was significantly higher than that in the adjacent non-cancerous tissues （2.181 ± 2.158 vs 0.931 ± 0.894, P = 0.0152）, and its over-expression was aggressively associated with lymph node metastasis （3.580 ± 2.487 vs 1.703 ± 0.758, P = 0.0003）, poorer cell differentiation （P = 0.0000） and advanced pathological stage （3.827± 2.673 vs 1.590 ± 0.735, P = 0.0001）. The patients were divided into high-expression and low-expression groups based on the median expression level of Bmi1 mRNA, and a shorter overall survival time in the former group was observed. Immunohistochemistry for Bmi1 oncoprotein showed diffusely positive, focally positive and negative expression in 44, 16 and 10 of 70 ESCC cases, respectively, compared with three, two and five of 10 adjacent non-cancerous cases （P = 0.027）. The positive rate of the oncoprotein in samples of histological grade Ⅲ was higher than that of grade Ⅱ（P = 0.031）, but its expression had no relation to the lymph node metastasis and pathological staging. In 70 ESCC samples, Bmi1 showed high intense expression in the cytoplasm and less or even no expression in the nucleus.CONCLUSION： Bmi1 was over-expressed in ESCC. Increased Bmi1 mRNA expression was significantly associated with ESCC progression, and the oncoprotein was largely distributed in the cytoplasm of tumor cells.