Serum matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase-1 in patients with malignant melanoma

Degradation of basement membranes and extracellular matrix is an essential step in cancer invasion and metastasis. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play key roles in this step. The present study was conducted to investigate the levels of MMP-3 and TIMP-1 in serum of patients with malignant melanoma and the relationship to tumor progression and known prognostic parameters. Seventy patients with cutaneous malignant melanoma were investigated. Serum samples were obtained on first admission before any adjuvant and metastatic treatment was given or follow-up of patients. Serum TIMP-1 and MMP-3 levels were determined by the solid-phase sandwich ELISA (Oncogene Science Inc.) method. The elevation of serum MMP-3 and TIMP-1 levels between the patients with malignant melanoma and healthy controls were not significantly different (p>0.05). The serum levels of MMP-3 were significantly different in males and females (p=0.001) and serum TIMP levels were influenced by age (p=0.047). Except for the ulceration status of the tumor, serum levels of MMP-3 and TIMP-1 were not related to the known prognostic factors such as tumor histology, localization, stage of the disease, Breslow thickness, Clark invasion, mitosis, TIL, and regression of tumor (p>0.05). In patients with ulceration positive, the serum levels of MMP-3 were higher (p=0.04) and TIMP-1 were lower (p=0.008) than those in patients without ulceration. No significant relationship was found between serum levels of MMP-3 and TIMP-1. In conclusion, these results suggest that neither of the serum levels of MMP-3 and TIMP-1 could be a good indicator of invasion and metastasis nor can be recommended as a tumor marker in the management of melanoma patients owing to lack of sensitivity and specificity. However, much research still continues in this field and exciting new knowledge will ultimately emerge.     

Serum vascular endothelial growth factor (VEGF) levels correlate with tumor VEGF and p53 overexpression in endocrine positive primary breast cancer

Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis, associated with unfavorable clinical characteristics in breast cancer. The aim of this study was to evaluate different angiogenic markers in endocrine-positive breast cancer patients. The authors analyzed serum and tumor samples from 71 patients with endocrine-positive operable primary breast cancer to determine the expression and the possible relationship between circulating serum VEGF levels, tumor VEGF expression, microvessel density (MVD), and other immunohistochemical parameters. Basal VEGF serum levels were significantly higher in breast cancer patients than in healthy controls. A significant correlation was observed between basal VEGF serum concentrations, microvessel density (p = 0.01) and p53 status (p = 0.004). Intratumoral VEGF expression was significantly associated with neoplastic embolization (p = 0.041) and circulating VEGF levels (p = 0.047). The results confirm that in primary endocrine-positive breast cancer serum VEGF levels are elevated and show a positive relationship with tumor VEGF and p53 overexpression.     

结肠癌组织MMP-2和MMP-9及VEGF表达临床意义的研究

目的:探讨结肠癌组织中基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)和血管内皮生长因子(VEGF)的表达及其与肿瘤进展的关系。方法:采用免疫组化SP法检测60例结肠癌组织中MMP-2、MMP-9和VEGF表达情况,并以25例正常结肠组织作为对照。结果:结肠癌组织中MMP-2、MMP-9和VEGF的阳性表达率分别为81.6%、78.3%和90.0%,高于正常结肠组织中的表达,两组比较差异有统计学意义,P<0.05。结肠癌组织中MMP-2、MMP-9和VEGF阳性表达与TNM分期和淋巴结转移相关,差异有统计学意义,P<0.05。结论:MMP-2、MMP-9和VEGF在结肠癌的发生发展和浸润转移中起到了重要的作用,可作为反映结肠癌进展的生物学指标。     

血管内皮生长因子、基质金属蛋白酶-2在直肠癌中的表达及其相关性

 【摘要】　目的　探讨血管内皮生长因子（VEGF）和基质金属蛋白酶－2（MMP-2）在直肠癌组织中的表达及其相关性， 并评价其在直肠癌进展中的作用及意义。方法　采用免疫组织化学SP法检测52例直肠癌患者癌组织中VEGF和MMP-2表达情况， 正常直肠组织12份作对照。结果　VEGF在直肠癌组织中阳性表达率为67.3 %， 与对照组比较差异有统计学意义（P＜0.01），且与淋巴结转移、Duke分期、侵袭深度和病理类型间差异有统计学意义（P＜0.01）。但在年龄和性别间差异无统计学意义（P＞0.05）。MMP-2蛋白在直肠癌组织中阳性表达率为71.2 %，显著高于对照组，差异有统计学意义（P＜0.01），与淋巴结转移和Duke分期间比较差异有统计学意义（P＜0.05）。VEGF、MMP-2之间存在显著的正相关（r＝0.478，P＜0.01）。结论　VEGF和MMP-2在直肠癌的发生、发展和浸润、转移过程中发挥着重要作用，可作为判断直肠癌转移潜能和预后的有效指标。     

Vascular endothelial growth factor (VEGF) upregulates BCL-2 and inhibits apoptosis in human and murine mammary adenocarcinoma cells.

Tumour progression is regulated by the balance of proliferation and apoptosis in the tumour cell population. To date, the role of vascular endothelial growth factor (VEGF) in tumour growth has been attributed to the induction of angiogenesis. VEGF has been shown to be a survival factor for endothelial cells, preventing apoptosis by inducing Bcl-2 expression. In both murine (4T1) and human (MDA-MB-231) metastatic mammary carcinoma cell lines, we found that VEGF upregulated Bcl-2 expression and anti-VEGF antibodies reduced Bcl-2 expression. These alterations in Bcl-2 expression were reflected by the levels of tumour cell apoptosis. VEGF resulted in reduced tumour cell apoptosis, whereas its inhibition with anti-VEGF neutralizing antibodies induced apoptosis directly in tumour cells. Therefore, in addition to its role in angiogenesis and vessel permeability, VEGF acts as a survival factor for tumour cells, inducing Bcl-2 expression and inhibiting tumour cell apoptosis.     

Expression of vascular endothelial growth factor, matrix metalloproteinase-9 and E-cadherin in the process of lymph node metastasis in oesophageal cancer.

Lymph node metastasis is a strong independent prognostic factor for oesophageal cancer. The expression of matrix metalloproteinases (MMPs) and reduction of E-cadherin correlate with lymph node metastasis of oesophageal cancer. We previously reported that the expression of vascular endothelial growth factor (VEGF) is associated with lymph node metastasis. This study was designed to determine whether VEGF, MMP-9 and E-cadherin expression is stable or changes in the process of lymph node metastasis of oesophageal cancer. Using immunohistochemistry, we detected VEGF, MMP-9 and E-cadherin expression in paraffin-embedded specimens of oesophageal squamous cell carcinoma. We classified 134 primary tumours and 174 nodal metastases using two different criteria: the absence [Group N(-)] or presence [Group N(+)] of nodal metastasis, and the stage of metastasis--Early Stage (cancer cells < 50% of lymph node) or Late Stage (> or = 50%)--and compared the expression among two groups and among two stages. The expression rates of Group N(-), Group N(+), Early Stage and Late Stage are as follows: VEGF (49%, 74%, 60%, 33%), MMP-9 (76%, 65%, 95%, 69%) and E-cadherin (49%, 24%, 55%, 38%). VEGF expression was down-regulated in Late Stage lymph node metastasis, while MMP-9 expression was elevated in Early Stage metastasis. E-cadherin expression is restored somewhat in Early Stage metastasis, but suppressed again in Late Stage metastasis. These data suggest that the expression of VEGF, MMP-9 and E-cadherin each change in the process of lymph node metastasis in oesophageal cancer, and that the patterns of change are different.     

基质金属蛋白酶-3和血管内皮生长因子在胃癌组织中的表达及意义

目的检测胃癌基质金属蛋白酶-3（MMP-3）和血管内皮生长因子（VEGF）的表达,探讨MMP-3和VEGF在不同临床病理特征中的意义,以期为临床工作提供理论支持。方法本实验以140例胃癌新鲜标本作为观察组,140例正常新鲜胃黏膜组织作为对照组,采用流式细胞技术,检测两组中MMP-3和VEGF表达的关系,观察MMP-3和VEGF的表达在不同临床特征中的差别。结果胃癌中MMP-3和VEGF的表达量明显高于对照组,MMP-3和VEGF的表达与胃癌的分化程度、浸润深度及淋巴结转移密切相关。MMP-3和VEGF在胃癌中的表达呈正相关性。结论胃癌中MMP-3和VEGF高表达,参与肿瘤的发生发展,联合检测MMP-3和VEGF的表达可能对判断胃癌的预后有一定价值。     

Correlation and expression of p53, HER-2, vascular endothelial growth factor (VEGF), and e-cadherin in a high-risk breast-cancer population

Background Many genetic traits common to aggressive breast carcinoma have been identified; yet little is known about the interrelationships of such traits during tumor development, especially in women prone to aggressive cancer. This study examined the expression of four biological markers associated with poor prognosis at each stage of breast cancer progression in primary tumors from women of lower economic status and assessed the relationship between these markers.Methods Archived primary breast tumors from 77 patients were assessed by immunohistochemical analysis for expression of human epidermal growth receptor 2 (HER-2), p53, vascular endothelial growth factor (VEGF), and e-cadherin, and the relationships between the expressions of these molecules were studied.Results Twenty-two (29%) patients had advanced (stage III or IV) disease. HER-2, VEGF, e-cadherin, and p53 signal were positive for 31 (40%), 58 (75%), 63 (82%), and 37 (48%) of patients, respectively. Among the markers tested, only p53 exhibited a significant association between expression and stage of the disease (P = 0.012). Expression of e-cadherin was positively associated with HER-2 overexpression (P = 0.004), and high levels of HER-2 occurred with strongly positive e-cadherin tumors. Marginally significant positive associations were observed between HER-2 and p53 signal (P = 0.06), and between disease stage and e-cadherin expression (P = 0.08).Conclusion The significant tendency toward expression of e-cadherin in conjunction with HER-2 overexpression in breast cancer is a novel finding. The association of p53 with more advanced stages of cancer emphasizes it as a key participant in metastatic processes in breast cancer.     

基质金属蛋白酶-2及基质金属蛋白酶-9的表达与胆囊癌转移、浸润的关系

目的 探讨胆囊癌组织中基质金属蛋白酶—2(MMP—2)及基质金属蛋白酶—9(MMP—9)的表达及其与胆囊癌浸润、转移的关系。方法 选取手术切除的胆囊癌标本42例及慢性胆囊炎标本15例，采用免疫组织化学的方法(SABC法)检测MMP—2及MMP—9蛋白的表达。并结合临床病理学指标进行统计学处理分析。结果 胆囊癌组织中MMP—2及MMP—9的阳性表达率显高于慢性胆囊炎。低分化及末分化组MMP—2的阳性率显性高于高分化组及中分化组。浆膜层浸润组、淋巴结转移组及肝脏转移组MMP—2的阳性率分别显性高于无浆膜层浸润组、无淋巴结转移组及无肝脏转移组。浆膜层浸润组及淋巴结转移组MMP—9的阳性率显高于无浆膜层浸润组及无淋巴结转移组。胆囊癌组织中MMP—2和MMP—9的表达有显的相关性。(P＜0．05)．二同时阳性表达组其浆膜层浸润、淋巴结和肝脏转移发生率显高于其中之一或均阴性组(P＜0．05)。结论 MMP—2的表达与胆囊癌的分化、浸润、淋巴结及肝脏转移密切相关。MMP—9的表达与胆囊癌浸润及淋巴结转移密切相关。MMP—2和MMP—9在胆囊癌组织中表达正相关，在胆囊癌的侵袭和转移过程中起协同作用。     

Pretreatment vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) serum levels in patients with metastatic non-small cell lung cancer (NSCLC)

PURPOSE: In the present study, we investigated the prognostic value of vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP)-9 serum levels in patients with metastatic non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: From September 1999 to June 2001, pretreatment serum levels of VEGF and MMP-9 were analysed in 194 patients of a randomized phase III trial with enzyme-linked immunoassays. RESULTS: Patients with a VEGF serum level higher than the median serum level (10,995 pg/ml) had a significantly shorter overall survival than those with a lower serum level (P=0.04). The MMP-9 serum level did not correlate with survival. In a multivariate Cox regression analysis, only the pretreatment serum level of VEGF, the Karnofsky performance status, and the presence of bone metastases were identified as independent prognostic factors. CONCLUSIONS: The pretreatment VEGF serum level was identified as independent prognostic factor in this study and may help to assess individual risk and treatment profiles in patients with metastatic NSCLC.     

血管内皮生长因子和基质金属蛋白酶-2在肝细胞肝癌中的表达及临床意义

目的研究血管内皮生长因子（VEGF）和基质金属蛋白酶-2（MMP-2）在肝细胞肝癌（以下简称肝癌）中的表达,及其与肝癌复发转移的关系。方法采用免疫组化方法检测90例肝癌手术切除标本的VEGF及MMP-2的表达情况。结果肝癌组织中VEGF与MMP-2的阳性率分别为76.7％与60.0％。MMP-2与VEGF的表达呈正相关（rs=0．32）,且两者均与复发转移正相关（rs=0.31,rs=0.32）。VEGF（-）组、VEGF（＋）组和VEGF（＋＋）组患者的2年无瘤生存率分别为71.4％、43．5％和30．4％（P〈0．01）;MMP-2（-）组和MMP-2（＋）组的2年无瘤生存率分别为66.7％和32．8％（P〈0.01）。多因素分析显示,肝癌组织中VEGF与MMP-2的表达水平、术前播散结节及镜下癌栓是肝癌复发转移的独立影响因素。结论肝癌组织中VEGF和MMP-2的表达水平结合临床病理特征（术前播散结节及镜下癌栓）,可评价肝癌患者术后复发转移的风险。     

VEGF与MMP-2在大肠癌组织的表达及相互关系的研究

目的 :检测血管内皮生长因子(vascularendothelialgrowthfactor ,VEGF)和基质金属蛋白酶 -2 (matrixmetalloproteinase 2 ,MMP 2 )在大肠癌组织中的表达及其与肿瘤进展的关系。方法 :应用免疫组化SP法检测了 18例正常大肠黏膜、18例大肠腺瘤样息肉和 5 1例大肠癌组织中VEGF、MMP 2的表达。结果 :大肠腺瘤样息肉、大肠癌中VEGF、MMP 2阳性表达率显著高于正常大肠黏膜组织 ,P <0 0 5。VEGF、MMP 2阳性表达率与大肠癌的淋巴结转移、Duke’s分期有关 ,P <0 0 5。VEGF、MMP 2在大肠癌中的表达有相关性 ,P<0 0 5。结论 :VEGF、MMP 2可能成为反映大肠癌进展的生物学指标及其治疗的理想靶点     

Expression of Vascular Endothelial Growth Factor-C and Vascular Endothelial Growth Factor Receptor-3 in Ovarian Epithelial Tumors

Objective： To explore the role of vascular endothelial growth factor-C （VEGF-C） in the process of angiogenesis, lymphangiogenesis and lymphatic metastasis in epithelial ovarian tumors. Methods： In situ hybridization and immunohistochemical staining for VEGF-C were performed in 30 epithelial ovarian carcinomas, 9 borderline tumors and 26 benign tumors. Endothelial cells were immunostained with anti-VEGFR-3 pAb and anti-CD31 mAb, and VEGFR-3 positive vessels and microvessel density （MVD） were assessed by image analysis. Results： VEGF-C mRNA and protein expression were detected in cytoplasm of carcinoma cells. VEGF-C mRNA and protein expression in ovarian epithelial carcinomas were significantly higher than those in borderline tumors and benign tumors （P〈0.05 or P〈0.01）. In ovarian epithelial carcinomas, VEGF-C protein expression, VEGFR-3 positive vessels and MVD were significantly higher in the cases of clinical stage Ⅲ-Ⅳ and with lymph node metastasis than those of clinical stage Ⅰ-Ⅱ and without lymph node metastasis respectively （P〈0.05 or P〈0.01）. VEGFR-3 positive vessels and MVD were significantly higher in VEGF-C protein positive tumors than negative tumors （P〈0.05）. VEGFR-3 positive vessels was significantly correlated with MVD（P〈0.01）. Conclusion： VEGF-C might play a role in lymphatic metastasis via lymphangiogenesis and angiogenesis in epithelial ovarian tumors, and VBEGF-C could be used as a biologic marker of metastasis in ovarian epithelial tumors.     

The constitutive level of vascular endothelial growth factor (VEGF) is more important than hypoxia-induced VEGF up-regulation in the angiogenesis of human melanoma xenografts

Angiogenesis of tumours might develop as a result of environmental conditions, such as hypoxia, and/or as a result of genetic alterations specific for tumour cells. The relative contributions of these mechanisms were investigated by comparing the in vivo expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) to the hypoxic fraction, the angiogenic potential and the vascular density of four human melanoma lines (A-07, D-12, R-18, U-25) grown intradermally in Balb/c nu/nu mice. VEGF expression, bFGF expression and expression of pimonidazole, a marker of hypoxic cells, were investigated by immunohistochemistry. An association between high VEGF and bFGF expression and high angiogenic potential was detected, suggesting an important role for VEGF/bFGF in the angiogenesis of melanomas. High VEGF/bFGF expression was also related to low hypoxic fraction and high vascular density. Thus, the constitutive, genetically determined level of VEGF was probably more important than hypoxia-induced upregulation in the angiogenesis of the melanoma xenografts.     

Correlations of oral tongue cancer invasion with matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF) expression

BACKGROUND AND OBJECTIVES: In oral tongue cancer, the degree of tumor invasion has a significant effect on the prognosis. We hypothesized that the destruction of extracellular matrix and neovascularization are related to tumor infiltration mechanism. By studying the tissues of early stage oral tongue cancer patients, we are intending to clarify the invasion-related factors. MATERIALS AND METHODS: To demonstrate the invasion process in early T-stage oral tongue cancer, the expressions of extracellular matrix destruction-related molecules (MMP-2, MMP-9) and neovascularization-related molecule (VEGF) were observed by immunohistochemical study. Also, staining of CD31 was done for quantification of neovascularization. We analyzed relationship between expression of each substances and tumor invasion depth, tumor free survival rates, and cervical lymph node metastasis rate. RESULTS: The expression rates of MMP-2, MMP-9, VEGF in 38 early oral cancer patients were 52.6%, 78.9%, and 52.6%, respectively. Significant correlation was found between the VEGF expression and microvessel density showed by CD31 immunohistochemical staining (P < 0.001). VEGF expressions were significantly related with tumor invasion depth (P = 0.002). The tumor-free survival rate of those patients with VEGF-positive tumors was significantly poorer than in those with VEGF-negative tumors (P = 0.019). CONCLUSIONS: These results indicate that VEGF is a useful marker for predicting the tumor invasion in patients with early tongue cancer.     

Circulating serum levels of angiogenic factors and vascular endothelial growth factor receptors 1 and 2 in melanoma patients

Angiogenesis is essential for tumor progression and metastasis; however, the angiogenesis regulators that are biologically relevant for melanoma are still unknown. In this study, we analyzed the circulating serum levels of potent angiogenic factors, including vascular endothelial growth factor (VEGF), angiogenin, transforming growth factor-beta1 and VEGF receptors, VEGFR1 and VEGFR2, in human melanoma patients. One hundred and fourteen patients with histopathologically verified cutaneous melanoma at different stages and 30 healthy controls were investigated. Serum levels of angiogenic factors and VEGF receptors were quantitatively analyzed by solid-phase enzyme-linked immunosorbent assay. The age of the patients (61 men and 53 women) ranged from 18 to 80 years; median age was 51 years. Serum transforming growth factor-beta1 (P < 0.001), VEGF (P = 0.006) and VEGFR1 (P = 0.007) levels were significantly higher in patients with melanoma than in the control group. No significant differences, however, exist in the serum angiogenin and VEGFR2 levels between melanoma patients and the controls. The positive correlations of elevated serum levels of transforming growth factor-beta1, VEGF and VEGFR1 with advanced stages of disease were found. Significant relationship was found only between serum levels of VEGF and VEGFR2. Elevated serum transforming growth factor-beta1 (P < 0.001) and VEGF levels (P = 0.0012) were found to be poor prognostic factors. Serum level of angiogenin and VEGF receptors, however, had no effect on survival. Our data suggest that the angiogenic serum factors, including VEGF, transforming growth factor-beta1 and VEGFR1, but not angiogenin and VEGFR2 were increased in melanoma patients, especially associated with advanced disease stages. The mechanism of VEGF regulation of angiogenesis may in part be due to enhanced proliferation of VEGFRs, especially VEGFR1.     

血管内皮生长因子在子宫颈癌组织中的表达及意义

目的　探讨血管内皮生长因子 (VEGF ,VEGFmRNA)在子宫颈癌组织中的表达及意义。方法　采用免疫组织化学技术SP法及原位杂交方法检测 33例宫颈癌 ,其中鳞癌 2 0例 ,腺癌 13例及 10例正常宫颈组织中VEGF及VEGFmRNA的表达情况。结果　宫颈癌组织切片中VEGF及VEGFmRNA皆有活跃表达。正常宫颈组织中VEGFmRNA呈阴性表达 ,VEGF蛋白可见弱阳性表达 ,两组比较差异有显著性 (P <0 .0 5 ) ;宫颈癌组织切片中的VEGFmRNA的表达与宫颈癌病理类型比较差异有显著性 (P <0 .0 5 ) ;而与宫颈癌临床分期及病理分化程度比较差异无显著性 (P >0 .0 5 )。结论　VEGF及VEGFmRNA的阳性表达在子宫颈癌的发生 ,发展中可能有着重要的意义。VEGF的检测对判断宫颈癌的病理类型有一定的临床价值。