Angiogenesis in cholangiocellular carcinoma: expression of vascular endothelial growth factor, angiopoietin-1/2, thrombospondin-1 and clinicopathological significance

Angiogenesis in cholangiocellular carcinoma (CCC) has rarely been investigated. The aim of this study was to determine the angiogenesis status of CCC and assess its relationship with angiogenic factors and clinicopathological characteristics. We examined 33 surgically resected CCC specimens. Tumor angiogenesis was assessed by microvessel density (MVD) using the anti-CD34 antibody, and the expression of VEGF, Ang-1, Ang-2, and TSP-1 was determined by immunohistochemistry. The mean (+/- SD) MVD was 87.2+/-52.6/mm2 (range, 0-229/mm2). A total of 75.6% cases were positive for VEGF expression, 36% for Ang-1, 57.6% for Ang-2 and 45.5% for TSP-1. VEGF and Ang-2 expression was associated with a significantly higher level of MVD (p=0.004 and 0.015, respectively). TSP-1 expression was associated with a significantly lower level of MVD (p=0.005) and a higher level of intrahepatic metastasis (46.7% vs. 5.6%, p=0.012). There was no significant correlation between VEGF, Ang-1, Ang-2, and TSP-1 expression and tumor size, capsule formation, infiltration of capsule, portal vein invasion, intrahepatic metastasis or CCC differentiation. There was no significant correlation between MVD levels, VEGF, Ang-1, Ang-2, and TSP-1 expression and postoperative survival. A considerable degree of angiogenesis, comparable to that of other solid tumors, was observed in CCC. VEGF and Ang-2 might play a proangiogenic role, and TSP-1 may play an inhibitory role in CCC. Although TSP-1 may increase intrahepatic CCC metastases, neither MVD levels nor the expression of VEGF, Ang-1, or Ang-2 was associated with clinicopathological factors and prognosis.     

肝细胞癌中NOS和VEGF的表达及其与肿瘤血管形成的关系

目的 研究Ⅱ、Ⅲ型一氧化氮合酶（ｉＮＯＳ、ｃＮＯＳ）和血管内皮生长因子（ＶＥＧＦ）在肝细胞癌中的表达及其与肿瘤血管形成的关系。方法 应用免疫组化方法检测７１例肝细胞癌患者手术切除石蜡包埋标本ｉＮＯＳ、ｅＮＯＳ、ＶＥＧＦ的表达,抗ＣＤ３４单克隆抗体显示血管内皮细胞。根据ＣＤ３４阳性的血管内皮细胞计数来测定肿瘤微血管密度（ＭＶＤ）。结果 ｉＮＩＳ、ｅＮＯＳ分别在８１．３％、８５．９％的肝癌中阳性表达,     

组织因子表达与胃癌微血管生成的关系及其临床意义

背景与目的组织因子(tissuefactor,TF)生理功能为凝血过程的启动者。近来发现TF参与多种恶性肿瘤微血管形成的过程。本研究旨在探讨胃癌组织中表达的组织因子与其微血管形成过程之间的关系和临床意义。方法应用免疫组织化学EnVisionTM法,检测了80例胃癌和20例正常胃组织标本中TF、VEGF(血管内皮生长因子)的表达情况和CD34单克隆抗体标记的MVD值(肿瘤微血管密度)。结果胃癌组织中TF和VEGF的阳性表达率分别为65.00%(52/80)和67.50%(54/80),MVD值为36.14±9.94;正常胃组织中TF和VEGF的阳性表达率分别为5.00%(1/20)和5.00%(1/20),MVD值为12.10±3.27。胃癌组织中TF的表达与VEGF的表达和MVD值之间均存在相关性(P<0.05),组织中TF表达越高,相应VEGF的表达越高,MVD值也越大。TF与患者的总生存期、TNM分期和肝脏转移状况有关。结论胃癌组织中异常表达增高的TF与胃癌微血管形成过程和患者预后有关。     

脑星形细胞瘤中TIP30、VEGF和CD34的表达及其相关性研究

目的探讨脑星形细胞瘤中TIP30、血管内皮生长因子（VEGF）、CD34的表达情况及其病理意义。方法采用免疫组织化学Elivision plus两步法,检测19例正常脑组织及71例星形细胞瘤组织中TIP30、VEGF及CD34的表达。结果TIP30在19例正常脑组织神经胶质细胞及神经元胞质内均呈阳性表达;71例星形细胞瘤组织中,TIP30总阳性表达率为33．80％,随星形细胞瘤分化程度的降低,TIP30的表达逐渐降低,Ⅱ、Ⅲ和Ⅳ级星形细胞瘤中TIP30阳性表达率分别为52％、34．78％和13．04％。高级别星形细胞瘤（Ⅲ级＋Ⅳ级）中TIP30的阳性表达率显著低于其在低级别星形细胞瘤（Ⅱ级）中的阳性表达率（x^2=5．71,P〈0．05）;VEGF及由CD34确定的MVD在星形细胞瘤中表达均高于正常脑组织,且随星形细胞瘤病理级别升高表达逐渐增高;星形细胞瘤中TIP30与VEGF强阳性表达呈负相关关系（r=-0．428,P〈0．05）。而TIP30与MVD无明显相关性（r=-0．065,P〉0．05）。结论TIP30在正常脑组织中阳性表达率高于其在星形细胞瘤中阳性表达率,且随星形细胞瘤病理级别升高表达显著下降;星形细胞瘤中,VEGF的表达与TIP30的表达呈明显的负相关关系。     

卵巢上皮癌组织微血管密度及血管形成相关分子的检测及预后价值

目的 探讨卵巢上皮癌组织微血管密度(MVD)、血管内皮生长因子(VEGF)、血小板反应素1(TSP1)和p53蛋白表达与患者预后的关系.方法 采用免疫组化法检测57例原发性卵巢上皮癌组织中VEGF、TSP1和p53蛋白的表达情况,用CD34免疫染色后计数MVD.对VEGF、TSP1、p53蛋白和MVD与患者复发及生存时间的关系进行回顾性分析.结果卵巢上皮癌组织中VEGF、TSP1和p53蛋白表达阳性率分别为70.2%(40/57)、47.4%(27/57)和61.4%(35/57),MVD为30.3±8.5,MVD、VEGF和TSP1与复发相关(P值分别为0.030、0.025和0.026).高MVD、VEGF和p53蛋白阳性患者的中位生存时间短于低MVD、VEGF和p53蛋白阴性者(P值分别为0.0187,0.010和0.005),MVD、VEGF和p53蛋白是影响预后的危险因素.TSP1是影响患者预后的保护因素,其阳性患者的中位生存时间长于阴性患者(P=0.042).多因素分析表明,MVD和p53蛋白是影响卵巢上皮癌预后的独立因素(P值分别为0.018和0.009).结论 VEGF、TSP1和p53蛋白可能参与了卵巢上皮癌的血管形成,MVD和p53是影响卵巢上皮癌预后的独立因素.     

CD147及相关因子在食管癌中的表达及意义研究

目的：探讨食管癌中细胞外基质金属蛋白酶诱导因子（EMMPRIN/CD147）在食管癌血管生成通路中的作用。方法随机抽取41例食管癌患者,其中淋巴结转移17例,应用免疫组化方法检测食管癌病理标本中CD147、血管内皮生长因子（VEGF）、血管内皮细胞生长因子受体2（VEGFR2）的表达及微血管密度（MVD）的计数。结果41例食管癌肿瘤组织中CD147、VEGF、VEGFR2阳性率分别为83%、54%及71%,明显高于正常组织。CD147、VEGFR2、MVD在淋巴结转移阳性与阴性患者之间存在显著差异（Z=-2.857,P=0.004;Z=-2.018,P=0.044;Z=-4.239,P﹤0.001）。食管造影下食管病变的长度与VEGF、CD147及MVD显著相关（r=0.486,P=0.01;r=0.588,P﹤0.001;r=0.604,P﹤0.001）。CD147与VEGF、VEGFR2、MVD之间均存在显著相关（r=0.976,P﹤0.001;r=0.588,P=0.01;r=0.604,P﹤0.001）。结论 CD147与VEGF、VEGFR2、MVD之间均存在显著相关,与食管病变长度密切相关,提示CD147在食管癌血管生成通路中有重要作用,且可能是通过调控VEGF通路实现的,但尚需进一步研究。     

乳腺癌血清VEGF水平与癌组织中VEGF和COX-2表达及微血管密度的关系

目的探讨乳腺癌患者血清血管内皮生长因子(sVEGF)水平与乳腺癌血管生成的关系。方法采用酶联免疫吸附试验(ELISA)检测68例乳腺癌、35例乳腺良性病变和20例健康女性的sVEGF水平,免疫组化S-P法检测相应乳腺癌组织中VEGF、环氧合酶-2(COX-2)及微血管密度(MVD)表达水平,并分析sVEGF水平与VEGF、COX-2及MVD表达的关系。结果(1)健康女性组、乳腺良性病变组和乳腺癌组sVEGF浓度中位数分别为105.93、150.82和306.51 pg/ml,乳腺癌组明显高于健康女性组。(2)乳腺癌组VEGF和COX-2表达阳性率分别为67.6%和44.1%,乳腺良性病变组VEGF和COX-2表达阳性率分别为42.9%和11.4%,两组间差异有统计学意义(P值分别为0.015和0.002)。(3)乳腺癌患者sVEGF水平与癌组织中VEGF、COX-2及MVD表达均呈正相关。(4)乳腺癌患者中,VEGF表达阳性组COX-2阳性率(65.21%)明显高于VEGF表达阴性组(18.18%); COX-2表达阳性组MVD(22.94±5.51)明显高于COX-2表达阴性组(10.30±4.42)。结论乳腺癌患者sVEGF水平明显增高于健康女性,并与癌组织中VEGF、COX-2及MVD表达呈正相关。     

Prognostic significance of vascular endothelial growth factor expression in human ovarian carcinoma

The influence of vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) on prognosis and the relationship between VEGF expression and MVD in ovarian carcinoma are not well defined. We studied VEGF expression in parallel with MVD by immunohistochemistry in 94 ovarian tumours (64 malignant, 13 borderline, and 17 benign) and correlated the results with the clinicopathologic prognostic factors of the disease to clarify their significance in this disease. Assessment of VEGF mRNA isoforms by RT-PCR was also performed. Of the malignant, borderline, and benign ovarian tumours respectively, two (3%), four (31%) and 16 (94%) were negative, 31 (48%), seven (54%) and one (6%) had low expressions, and 31 (48%), two (15%) and none (0%) had high expressions of VEGF. There were significant associations between the VEGF expression and disease stage (P= 0.002), histologic grade (P= 0.0004), and patient outcome (P= 0.0002). MVD did not correlate significantly with the clinicopathologic parameters. Likewise, no correlation was found between MVD and VEGF expression. The survival of patients with high VEGF expression was significantly worse than that of patients with low and negative VEGF expression (P = 0.0004). Multivariate analysis revealed that disease stage and VEGF expression were significant and independent prognostic indicators of overall survival time (P = 0.008 and P = 0.006 respectively). These findings suggest that in conjunction with the established clinicopathologic prognostic parameters of ovarian carcinoma, VEGF expression may enhance the predictability of patients at high risk for tumour progression who are potential candidates for further aggressive therapy.     

The natural progression of microvasculature in primary tumor and lymph node metastases in a breast carcinoma model: relationship between microvessel density, vascular endothelial growth factor expression, and metastatic invasion

The natural course of tumor microvascularity in rat MTLn3 mammary adenocarcinomas was studied. The relationship between microvessel density (MVD), vascular endothelial growth factor (VEGF) expression, and histopathology was compared in primary and metastatic axillary (ALN) and inguinal lymph node (ILN) tumors over 5-6 tumor doublings. Excised tumors were examined for (i) MVD assessed by immunostaining with anti-CD31 antibody, (ii) VEGF expression assessed by immunostaining with anti-VEGF antibody, and (iii) histopathologic extent of metastatic lymph node invasion. MVD and VEGF scores rose asymptotically with increasing tumor weight in both primary and metastatic tumors. The MVD saturation level was significantly greater for primary tumors (MVD = 22) than for ALNs or ILNs (MVD = 14). Maximal VEGF score was not statistically different between the three kinds of tumors, however the rate of rise in VEGF expression was different. Near-maximal VEGF expression occurred early in tumor growth, preceding microvessel development. Both MVD and VEGF expression in lymph nodes were proportional to the pathology score characterizing increasing metastatic invasion. LNMs limited to the subcapsular sinus had the lowest MVD, indicating an ability to survive without significant vasculature. These findings underscore the differences in angiogenesis between primary tumors and LNMs and have implications for therapy of metastatic cancer.     

CD147, MMP-2, MMP-9 and MVD-CD34 are significant predictors of recurrence after liver transplantation in hepatocellular carcinoma patients

OBJECTIVE: The prognostic role of tumor biological markers (biomarkers) in predicting recurrence of hepatocellular carcinoma (HCC) was investigated in this study, the results of which may help to select appropriate candidates for liver transplantation (LT). METHODS: Tissue samples from 82 HCC patients with cirrhosis who had undergone LT were immunohistochemically stained with antibodies of anti-CD147, anti-matrix metalloproteinases-2 (MMP-2), MMP-9 and anti-vascular endothelial growth factor (VEGF). Tumor microvessel density (MVD) was evaluated by using CD34. Multivariate Cox regression analysis was performed to identify the relevant prognostic factors. RESULTS: A significant correlation was found between the expression of CD147, VEGF, MMP-2, MMP-9 and MVD-CD34 in HCC. Tumor CD147 expression (P < 0.0001), tumor MVD-CD34 (P < 0.0001), MMP-9 in stromal compartment (P = 0.0257) and tumor VEGF expression (P = 0.0335) were significantly associated with the recurrence in HCC patients after LT. Univariate analysis showed that strong CD147 expression and high MVD-CD34 were significantly associated with poor tumor recurrence-free survival after LT (P < 0.0001). Multivariate analysis indicated that CD147 (P = 0.0001), MVD-CD34 (P = 0.0118), MMP-2 (P = 0.0312) and MMP-9 (P = 0.0280) in stromal compartment were all significant predictors in predicting HCC recurrence, while VEGF, MMP-2 and MMP-9 in tumor compartment were not significantly associated with poor prognosis. CONCLUSIONS: The tumor biomarkers CD147 and MVD-CD34 are more feasible markers for rational selection of LT candidates with HCC. MMP-9 and MMP-2 expression in stromal compartment, combined with pTNM tumor grade, may be helpful in predicting poor prognosis in HCC patients after LT.     

Expression of CD34 in gastric cancer and its correlation with histology, stage, proliferation activity, p53 expression and apoptotic index

The formation of new blood vessels is essential for tumor growth and progression. Until today there are only few studies of the immunohistochemical assessment of angiogenesis in gastric cancer by the evaluation of the expression of CD34 antigen. The aim of this study was to analyze the relationship between microvessel density (MVD) expressed as the mean count of CD34 immunostained vessels and clinicopathologic features of gastric tumors (the histological type according to the Lauren classification, tumor grade G; presence of lymph node metastases N; depth of tumor invasion; stage of disease (UICC-AJCC 1988 1992), p53 expression, tumor cell proliferative activity described as the Ki67 labelling index and apoptotic index of tumor cells TUNEL method). We assessed formalin-fixed, paraffin-embedded tissue samples obtained during potentially radical gastrectomy from 58 patients with primary gastric adenocarcinoma. The representative tissue blocks from each tumor were used for the immunohistochemical assay and examined by two pathologists independently. MVD was counted in five tumor areas of the most intensive neovascularization (x 200 field by light microscopy) and the mean counts were recorded. The mean MVD (CD34 expression value+/-SD) in this study was 43,15+/-19,8 per x 200 field. The study demonstrated the statistically significant correlation between MVD and two main histological parameters: tumor grading (p < 0.001) and tumor histological type according to Lauren s classification (p<0.05). In well and moderately differentiated tumors (G1/2) MVD was significantly lower in comparison to the group of poorly differentiated cancer G3 (mean value: 31,62 vs. 49,89). MVD was higher in diffuse type of gastric cancer comparing to intestinal type (50.05+/-19,03 vs. 39.17+/-20,09). However, the authors failed to find a significant correlation between MVD and other investigated histopathological features in malignant gastric tumors. The close relationship between CD34 immunostaining, gastric cancer tumor vascularity and main histological parameters was shown in this study. It can be stated that analysis of expression of angiogenesis in gastric cancer may be helpful for better estimation of hematogenous recurrence and the selection of the group of patients for adjuvant antiangiogenic treatment.     

Expression of caveolin-1 in mucoepidermoid carcinoma of the salivary glands: correlation with vascular endothelial growth factor, microvessel density, and clinical outcome

BACKGROUND: Caveolin-1, which has been proposed as a candidate tumor suppressor, plays a regulatory role in several signaling pathways. The importance of caveolin-1 in endothelial cells in angiogenesis has been confirmed. The clinicopathologic significance of caveolin-1 expression and its correlation with angiogenesis remains unknown in mucoepidermoid carcinoma (MEC) of the salivary glands. METHODS: Based on an immunohistochemical study, the expression levels of caveolin-1 and vascular endothelial growth factor (VEGF) and the intratumoral microvessel density (MVD) (labeled by CD34) in 75 patients with MEC were investigated, and correlations with clinicopathologic variables were evaluated statistically. RESULTS: The expression rates of both caveolin-1 and VEGF were 54.7% (41 of 75 tumors). MVD varied from 9 to 56 (24.45 +/- 10.72)/x 200. Caveolin-1 expression was correlated inversely with duration of tumor, clinical stage, histologic grade, and MVD (P = .027, P = .011, P = .04, and P = .025; respectively). VEGF expression was associated positively with MVD (P = .000). Advanced clinical stage, higher grade, and tumors that originated from minor salivary glands exhibited higher MVD (P = .029, P = .002, and P = .008, respectively). The presence of clinical symptoms, male gender, advanced clinical stage, higher grade, increased MVD, and down-regulated caveolin-1 were correlated significantly with the development of recurrent disease, as indicated by a shorter disease-free interval (P < .05). Both univariate and multivariate analyses indicated that clinical stage, histologic grade, and MVD were independent prognostic factors (P < .05). The presence of clinical symptoms and the down-regulation of caveolin-1 were identified as negative prognostic predictors in the univariate analysis (P < .05) but did not achieve significance in the multivariate analysis (P > .05). CONCLUSIONS: The current results suggest that caveolin-1 may function as a tumor suppressor in MEC of the salivary glands. Reduced expression of caveolin-1 and increased MVD may indicate a poor prognosis for certain patients.     

Ang-2、CD105和CD34在脑胶质瘤中的表达及生物学意义

目的：探讨促血管生成素-2（Ang-2）、CD105和CD34在脑胶质瘤中的表达及生物学意义。方法：应用免疫组化sP法检测45例胶质瘤和10例正常脑组织中Ang-2蛋白的表达及CDl05和CD34标记的MVD，并分析它们与胶质瘤临床病理因素的关系。结果：Ang-2在胶质瘤中的阳性表达率为62．2％，显著高于正常脑组织（P〈0．05）。胶质瘤组织中，CD105、CD34标记的MVD均明显高于正常脑组织。Ang-2表达的阳性标记指数及CD105标记的MVD与胶质瘤病理分级密切相关（P〈0．01），CD34标记的MVD与胶质瘤病理分级无明显相关（P〉0．05）。Ang-2与CD105、Ang-2与CD34均呈协同表达，CD105在胶质瘤中表达的特异性较CD34高。结论：Ang-2、CD105和CD34表达在促进胶质瘤的恶性演进中发挥重要作用；CD105较CD34更能准确反映肿瘤血管内皮细胞的增殖状态。     

Expression of vascular endothelial growth factor and microvessel density in head and neck tumorigenesis.

Angiogenesis is a fundamental process in tumor growth and metastasis, and its significance and that of vascular endothelial growth factor (VEGF) expression as prognostic indicators have been documented for various types of human tumors. However, the mechanisms responsible for angiogenesis in head and neck squamous cell carcinoma are not well defined. To examine the relationship between angiogenesis and the phenotypic progressions of head and neck tumorigenesis, we used immunohistochemistry to analyze VEGF expression and microvessel density in 70 paraffin-embedded specimens that contained adjacent normal epithelium, premalignant lesions, or both from 57 patients with head and neck squamous cell carcinoma. Ten samples of normal oral mucosa were obtained from people who did not smoke or drink alcohol and included in the analysis as normal controls. Microvessel density was evaluated by averaging 10 microscopic fields (x400) in a defined area of each specimen. The degree of VEGF expression was assessed on a cell-by-cell basis in 10 microscopic fields (x200) in a defined area on a scale ranging from 0 (no expression) to 3+ (highest level of expression). In addition, the weighted mean index of VEGF expression was calculated. The mean +/- SD weighted mean index of VEGF expression in normal control epithelium (1.10 +/- 0.38, n = 10) was higher than it was in adjacent normal epithelium (0.82 +/- 0.27, n = 13; P = 0.04). VEGF expression decreased as samples ranged from normal adjacent epithelium to hyperplasia (0.78 +/- 0.28, n = 21), mild dysplasia (0.70 +/- 0.29, n = 28), moderate dysplasia (0.67 +/- 0.29, n = 11), severe dysplasia (0.51 +/- 0.39, n = 6), and squamous cell carcinoma (0.20 +/- 0.27, n = 70; overall P = 0.0001). VEGF expression was two times lower in cases with nodal disease (0.17 +/- 0.26, n = 29) than it was in nonnodal disease (0.32 +/- 0.29, n = 16; P = 0.02). Microvessel density showed no significant difference from adjacent normal epithelium premalignant lesions to cancer. In tumor, no correlation was seen between VEGF expression or microvessel density and differentiation, primary tumor site, T stage, or smoking status. These findings indicate that VEGF expression is down-regulated during head and neck tumorigenesis. However, further studies are required to better understand the mechanism of VEGF down-regulation in head and neck tumorigenesis.     

结直肠肿瘤微血管计数及血管内皮细胞生长因子表达

目的 探讨血管生成与结直肠肿瘤的发生,发展关系,评估微血管计数（ＭＶＤ值）及血管内皮细胞生长因子（ＶＥＧＦ）表达与结直肠肿瘤预后的相关性。方法 应用免疫组化法回顾性地研究了３２例结直肠肿瘤石蜡包埋的病理组织。结果 正常粘膜,腺瘤,癌组织的ＭＶＤ值递增。不同病理状态下的结直肠癌ＭＶＤ值有差异,ＶＥＧＦ阴性组ＭＶＤ值低于ＶＥＧＦ阳性组,低ＭＶＤ值主ＶＥＧＦ阴性组生存率高于高ＭＶＤ值组及ＶＥＧＦ阳性组。     

Vascular endothelial growth factor/KDR activated microvessel density versus CD31 standard microvessel density in non-small cell lung cancer

Vascular endothelial growth factor (VEGF) is an important angiogenic factor, linked to poor outcome in human malignancies including non-small cell lung carcinoma (NSCLC). We used the 11B5 monoclonal antibody recognizing the VEGF/KDR complex (R. A. Brekken et al., Cancer Res., 58: 1952-1959, 1998) to assess the VEGF expression in cancer cells and the VEGF/KDR activated microvessel density (aMVD) in early operable NSCLC. The JC70 anti-CD31 monoclonal antibody was used to assess the standard MVD (sMVD). The aMVD was significantly higher in the invading front of the tumors and in the normal lung adjacent to the tumors as compared with normal lung distant to the tumor or to inner tumor areas (P < 0.0002). The sMVD was higher in the normal lung and decreased from the invading front to inner tumor areas (P < 0.0001). However, the vascular activation (aMVD:sMVD) was 4-6 times higher in the tumor areas as compared with lung from normal individuals (36-58% versus 9%; P < 0.0001). Fibroblast 11B5 reactivity, noted in 25% of cases, correlated with high aMVD and sMVD in the inner tumor areas. Multivariate analysis showed that aMVD was the most potent and independent prognostic factor (P = 0.001; t-ratio, 3.28). It is concluded that intense VEGF/KDR angiogenic pathway activation is a tumor-specific feature in more than 50% of NSCLC cases and is associated with poor postoperative outcome. Clinical trials involving targeting of the VEGF/KDR-positive vasculature with specific antibodies, such as 11B5, are, therefore, encouraged.     

尿纤溶酶原激活物及血管内皮生长因子在食管癌中的表达及对肿瘤血管形成的影响

目的探讨尿纤溶酶原激活物（uPA）、血管内皮生长因子（VEGF）在食管癌中的表达及对肿瘤血管生成的影响。方法采用免疫组织化学sP法检测正常食管黏膜上皮组织（18例）及食管癌组织（68例）中uPA、VEGF的表达,检测CD。用以标记肿瘤微血管密度（MVD）,根据MVD均值分为高、低MVD组,分析食管癌uPA、VEGF的表达和临床病理特征的关系及对肿瘤血管形成的影响。结果uPA蛋白在正常食管黏膜上皮组织、食管癌组织中的阳性率分别为27．8％（5／18）和70．6％（48／68）,差异有统计学意义（X^2=11．63,P〈0．05）;VEGF蛋白在正常食管黏膜上皮组织、食管癌组织中的阳性率分别为22．2％（4／18）和63．2％（43／68）,差异有统计学意义（X^2=9．78,P〈0．05）。食管癌组织中uPA与VEGF表达有一致性（X^2=9．72,P〈0．05）。MVD平均为42．38±11．62,高MVD组uPA、VEGF蛋白表达显著高于低MVD组（X^2值分别为6．13和10．12,均P〈0．05）。uPA、VEGF蛋白表达与年龄、性别、病理类型无关（均P〉0．05）,均与临床病理分期、分化程度和淋巴结转移相关（P〈0．05）。结论食管癌组织中uPA、VEGF蛋白高表达,可能促进肿瘤血管形成,提示预后不良。     

Expressions of nuclear factor kappaB, inducible nitric oxide synthase, and vascular endothelial growth factor in adenoid cystic carcinoma of salivary glands: correlations with the angiogenesis and clinical outcome.

OBJECTIVE: To evaluate the expressions of nuclear factor kappaB (NF-kappaB p65), inducible nitric oxide synthase enzyme (iNOS), and vascular endothelial growth factor (VEGF) in relation to angiogenesis (microvessel density, MVD) and clinical outcomes in adenoid cystic carcinoma (ACC) of salivary glands. METHODS: Immunohistochemical staining was used to quantify the protein expression levels of NF-kappaB p65, iNOS, and VEGF in 80 surgically resected ACCs and 20 normal salivary tissues. In all cases of ACCs, MVD was evaluated by counting CD34-reactive endothelial cells or endothelial cell clusters. RESULTS: The nuclear localization of NF-kappaB p65 was only detected in ACC cells. Both iNOS and VEGF staining activities in ACCs were more significant than those in normal gland tissues (P < 0.01). MVD had significant correlations with NF-kappaB p65, iNOS, and VEGF expressions (P < 0.01). In three histologic types of ACCs, the NF-kappaB, iNOS, VEGF expressions, and MVD were significantly higher in solid type than in cribriform and tubular types (P < 0.01). The NF-kappaB, iNOS, VEGF expressions, and MVD were significantly correlated with clinical stage, tumor size, vascular invasion, recurrence, and metastasis (P < 0.05). Multivariate analysis showed NF-kappaB, iNOS and VEGF expression, MVD, solid histotype, and perineural invasion had an independent prognostic effect on overall survival. CONCLUSION: The expressions of NF-kappaB p65, iNOS, and VEGF were related with MVD. Clinical outcomes raised the possibility that the overexpression of these cytokines might contribute to tumor angiogenesis and have prognostic value in ACCs.     

P53蛋白、血管内皮生长因子等生物分子指标与鼻咽癌放射敏感性的关系

背景与目的:临床上常见鼻咽癌患者虽属于同一分期,并采用相同的治疗技术和治疗剂量,却有20%～40%患者在5年内出现射野内复发,这主要与肿瘤细胞的放射治疗内在敏感性个体差异有关。本研究对鼻咽癌细胞增殖、凋亡以及肿瘤血管生成、淋巴管生成等状况与放疗敏感性的关系作一初步探讨。方法:对放疗敏感及放疗抗拒的鼻咽癌患者活检标本各60例行免疫组化法检测P53、血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)、Survivin、VEGF-C、Ki67蛋白及微血管密度(microvasculardensity,MVD)表达,结合临床资料,分析它们之间的相关性。结果:P53、VEGF、Survivin、VEGF-C、Ki67蛋白阳性率分别为65.0%、57.5%、60.8%、42.5%、57.5%,放射敏感组及放射抗拒组P53、VEGF、MVD表达分别为(25.97±21.26)%、(18.50±19.86)%、32.65±19.61及(37.85±28.67)%、(30.83±23.94)%、41.95±16.97,两组间差异有显著性(P<0.05),Survivin、VEGF-C、Ki67蛋白阳性率两组间差异无显著性(P>0.05)。P53、VEGF、MVD三者间显著相关。结论:P53、VEGF蛋白及MVD在放疗抗拒组比放疗敏感组明显增高,有可能作为放疗前评估鼻咽癌内在放射敏感性的指标。     

In oesophageal squamous cell carcinoma vascular endothelial growth factor is associated with p53 mutation, advanced stage and poor prognosis.

Vascular endothelial growth factor (VEGF) affects malignant tumours by promoting angiogenesis. The tumour-suppressor gene p53 has been thought to regulate VEGF. We investigated the effect of VEGF on oesophageal carcinoma and the connection between VEGF and p53. One hundred and nine resected oesophageal squamous cell carcinomas were examined. VEGF expression was analysed by immunohistochemical staining. Sixty-five tumours (59.6%, 65 out of 109) were classified as VEGF positive. A significant correlation was found between the VEGF expression and both the depth of invasion (P = 0.0001) and lymph node metastasis (P < 0.0001). With regard to p53, we compared the expression of VEGF with the mutation of p53, examined using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and direct sequencing in tumour samples obtained from 36 patients who we have reported previously. The VEGF expression was significantly correlated to p53 mutation (P = 0.0291). To evaluate the angiogenesis, microvascular density (MVD) was counted, and endothelial cells were stained immunohistochemically using anti-CD34 monoclonal antibody against 29 cases with invasion limited to the submucosal layer. The average MVD had a tendency to correlate to VEGF expression (P = 0.1626). The prognoses of patients with VEGF-positive primary tumours were significantly worse than for those with VEGF-negative primary tumours (P = 0.0077). We have assumed that VEGF contributes to aggressive characteristics in oesophageal carcinomas and that VEGF expression might be affected by p53 status.