Expression of nerve growth factor and hypoxia inducible factor-1α and its correlation with angiogenesis in non-small cell lung cancer

Summary： In order to investigate the expression of nerve growth factor （NGF） and hypoxia inducible factor-1α （HIF-1α） and its correlation with angiogenesis in non-small cell lung cancer （NSCLC）, paraffin-embedded tissue blocks from 20 patients with NSCLC were examined. Twenty corresponding para-cancerous lung tissue specimens were obtained to serve as a control. The expression of NGF, HIF-1α, and vascular endothelial growth factor （VEGF） in the NSCLC tissues was detected by using immunohistochemistry. The microvascular density （MVD） was determined by CD31 staining. The resuits showed that the expression levels ofNGF, HIF-1α and VEGF in the NSCLC tissues were remarkably higher than those in the para-cancerous lung tissues （P〈0.05）. There was significant difference in the MVD between the NSCLC tissues （9.19±1.43） and para-cancerous lung tissues （2.23±1.19） （P〈0.05）. There were positive correlations between NGF and VEGF, between HIF-1α and VEGF, and between NGF and HIF-1α in NSCLC tissues, with the spearman correlation coefficient being 0.588, 0.519 and 0.588, respectively. In NSCLC tissues, the MVD had a positive correlation with the three factors （P〈0.05）. Theses results suggest that NGF and HIF-1α are synergically involved in the angiogenesis of NSCLC.     

Over-expression of hypoxia-inducible factor 1 alpha increases angiogenesis of LNCaP cells

Objective:To evaluate the effect of HIF-1 α over-expression on angiogenesis in human prostate cancer cells. Methods:LNCaP cells(a human prostate cancer cell line) were transfected with the recombinant plasmid pcDNA3.1(-)-HIF-1α with Lipofectamine 2000 system. The positive clones were selected by G418 being further confirmed by Western blot and immunofluorescence. The expression levels of VEGF, iNOS and Ang- Ⅱ were determined. Results:The expression of HIF-1α in the LNCaP/HIF1α cells was significantly increased in transfected cells, which induced the up-regulation of VEGF, iNOS, whereas Ang- Ⅱ expression remained un- changed. Conclusion :Over-expression of HIF-1α can induce angiogenesis proteins and may improve the angiogenesis potency of prostate cancer.     

Expression of pigment epithelium-derived factor in bladder tumour is correlated with interleukin-8 yet not with interleukin-1α

Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In this study the expression of PEDF, interleukin-1α (IL-1α) and -8 (IL-8) in bladder tumours was investigated. Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples. Expression change of the factors was compared with clinicopathological parameters. Correlations between PEDF, IL-1α and IL-8 were analyzed. None of the factors was in relation to gender, tumour occurrence, and size or onset pattern. PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression. PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping. PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049, r=-0.578) as well as in tumour grouping (P=0.033, r=-0.276). Deranged expressional change of PEDF, IL-1α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.     

Expression and Clinical Implication of HIF-1α and VEGF-C in Non-small Cell Lung Cancer

To study the expression and implication of HIF-1α and VEGF-C in non-small cell lung cancer （NSCLC） and its relationship with clinical pathological features of NSCLC, immunohisto-chemical SP was used to detect the expression of HIF-1α and VEGF-C proteins in 48 NSCLC tissues and the same para-cancerous tissues. The positive rates of HIF-1α and VEGF-C were 70.8% （34/48） and 68.8% （33/48） respectively. The expression of HIF-1α protein was detected in a significantly greater proportion in NSCLC carcinoma tissues than that in para-cancerous tissues （12.5% and 16.7%, P〈0.05）. The positive rates of HIF-1α and VEGF-C were correlated with lymph node metastasis and TNM stage. No relationship was found between the two factors and age, sex, pathological subtypes and histological grades. The positive rates between HIF-1α and VEGF-C were correlated （P〈0.05）. HIF-1α and VEGF-C were over-expressed in NSCLC. They may be involved in the carcinogenesis of NSCLC, and play an important role in invasion and metastasis of NSCLC. HIF-1α and VEGF-C work synergically in the process of NSCLC.     

Effects of TNF-α and curcumin on the expression of VEGF in Raji and U937 cells and on angiogenesis in ECV304 cells

Background To better understand the possibilities of antiangiogenic tumor therapy and to assess possible side effects, we investigated the effect of tumour necrosis factor （TNF）-α and curcumin on the expression of vascular endothelial growth factor （VEGF） in U937 and Raji cell lines and their effect on angiogenesis in a human umbilical vein endothelial cell （HUVECs）-derived cell line （ECV304）, and also the relationship between Notchl and VEGF. The aim of this study was to elucidate potential mechanisms controlling tumor neovascularization. Methods VEGF secreted by U937 and Raji cell lines was determined by ELISA. Angiogenesis was tested by network formation of endothelial cells on Matrigel. Levels of VEGF mRNA in U937 and Raji cells and Notchl mRNA levels in EV304 cells were determined by RT-PCR. Results Secretion of VEGF by U937 and Raji cells was increased by TNF-α treatment and suppressed by curcumin （P 〈 0. 01 ）. The mRNA expression of VEGF165 and VEGF121 （containing 165 and 121 amino acid residues, respectively） were detected in any fractions. TNF-α augmented the expression of VEGF165 and VEGF121 mRNA and curcumin reduced the expression （P 〈0. 01 ）. No networks or cords formed in control and curcumin groups. There was tube formation on matrigel in the supernatants of the Raji culture group and the supernatants groups treated by VEGF group and TNF-α in Raji cell. Notch1 mRNA was detected but there was no significant change in the VEGF group compared with control （P 〉 0. 05）. Conclusions Expressions of VEGF mRNA in U937 and Raji cells were increased by TNF-α and suppressed by curcumin. VEGF and TNF-α can induce angiogenesis, and curcumin can inhibit angiogenesis in ECV304 cells.     

Correlation between the Expression of Aquaporin 1 and Hypoxia-inducible Factor 1 in Breast Cancer Tissues

The correlation between aquaporin 1 （AQP1） and hypoxia-inducible factor 1 （HIF 1） in breast cancer tissues was preliminarily studied. In 155 cases of breast cancer, the expression levels of AQP1 were detected by immunohistochemisty in HIFl-positive group or HIFl-negative group, and the correlation between AQP1 and HIF1 was analyzed. The overexpression of AQP1 and HIF1 were observed in 155 cases of breast cancer tissues. The expression level of AQP1 in HIFl-positive group was significantly higher than that in HIFl-negative group. The positive expression rate of AQP1 was 296.55±24.67 and 168.37±37.53 in HIFl-positive group and HIFl-negative group respectively with the difference being very significant between them （P〈0.001）. It was concluded that AQP1 was overexpressed in the HIFl-positive group and there were some correlations between AQP1 and HIF1, suggesting they interact each other and regulate the oncogenesis of breast cancer.     

Correlation between the expression of thrombospondin-1 and neovascularization in mucoepidermoid carcinoma

Background Researchers have recently demonstrated that thrombospondin-1 （TSP-1） has an important function in regulating neovascularization. Whether it inhibits or accelerates neovascularization, however, is still controversial. We found few reports about the correlation between TSP-1 and vascularization in mucoepidermoid carcinoma. In this research, the distribution and expression of TSP-1 in mucoepidermoid carcinoma were investigated. We also analyzed （1） the correlation between the expression of TSP-1 and microvessel density （MVD）, as an indicator of neovascularization activity, and （2） the effect of TSP-1 on neovascularization and tumor growth in the subcutaneous xenotransplanted model of mucoepidermoid carcinoma. Method （1） The sites and intensity of expression of TSP-1 and the MVD were analyzed in 45 cases of mucoepidermoid carcinoma after surgery by the method of streptavidin-peroxidase （SP） immunohistochemistry; and （2） recombinant human thrombospondin-1 （rhTSP-1） was injected twice a week for five consecutive weeks around the tumor in the subcutaneous xenotransplanted tumor model of mucoepidermoid carcinoma in nude mice. Each week, the tumor size was measured, in order to draw the growth curve of the xenotransplanted tumor model of mucoepidermoid carcinoma, and MVD was measured. Results （1） The positive expression of TSP-1 protein was 57.78% （26/45）. Most positive staining for TSP-1 was found in the cytoplasm of the cancer cells, while some staining occurred in the extracellular matrix. The mean MVD in 45 cases of m ucoepidermoid carcinoma was 58.17±19.77 per 100 visual fields. Tumors with a high expression of TSP-1 showed a low MVD value, and the TSP-1 immunocompetence and microvessel density showed a significant negative correlation （rs=-0.947, P 〈0.001）. （2） The xenotransplanted tumors with the injection doses of 1.25, 0.75 and 0.25 ug/ml respectively were 36.97%, 53.36% and 73.61% of the size of the control group （（451±92）, （651±113）, （8     

Correlation of vascular endothelial growth factor and CD105-microvascular density in primary pterygium

The relationship between the expression of vascular endothelial growth factor(VEGF) and microvascular density(MVD) marked by CD105(CD105-MVD),and that between CD105-MVD and the clinicopathological characteristics of primary pterygium were investigated.The streptavidin-biotin complex(SABC) immunohistochemical staining in paraffin-embedded tissues was used to detect the expression of VEGF in 23 cases of primary pterygia and 7 normal conjunctival specimens.The antibody against CD105 was used to display vascular endothelial cells,and MVD was examined by counting the CD105-positive vascular endothelial cells.The correlations of VEGF and CD105-MVD,and those of CD105-MVD and clinicopathological data were analyzed by using SPSS 12.0.The expression of VEGF was significantly increased in epithelia(P=0.000),endothelia and stroma cells(P=0.005) in primary pterygia as compared with normal conjunctivae.The CD105-MVD in pterygia(mean 19.22±6.68) was higher than that in normal conjunctivae(mean 4.00±2.15,P=0.000).MVD in pterygia was significantly associated with the Tan classification(P=0.000) and the VEGF expression level in the stroma(P=0.020),but not with sex(P=0.61),age(P=0.150) or the VEGF expression level in the epithelia(P=0.518).Our results suggest that over-expression of VEGF and high CD105-MVD in primary pterygium may contribute to the progression by increasing angiogenesis and growth of primary pterygium.     

siRNA沉默EPAS1基因抑制裸鼠胰腺癌模型血管生成

目的通过RNA干扰技术抑制缺氧诱导因子2（EPAS1）表达,从而抑制肿瘤新生血管形成来治疗胰腺癌。方法建立人胰腺癌裸鼠皮下移植瘤模型,分为实验组、阴性组和空白组,分别注射rAAV2-EPAS1-siRNA、rAAV2-EGFP和生理盐水。测量肿瘤体积和质量,免疫组化检测肿瘤EPAS1、血管内皮生长因子（VEGF）蛋白表达及微血管密度（MVD）值。结果与阴性组和空白组比较,实验组皮下瘤体积和质量降低,EPAS1、VEGF蛋白表达受抑制,微血管密度减小。结论siRNA沉默EPAS1基因能抑制肿瘤血管生成和胰腺癌生长。     

HIF-1α、VEGF在子宫内膜癌中的表达及其与肿瘤血管生成的关系

目的 研究子宫内膜癌组织中乏氧诱导因子-1α(HIF-1α)与血管内皮生长因子(VEGF)的表达情况及其与肿瘤血管生成的关系.方法 选取2012年2月至2015年12月河北工程大学附属医院收治的48例子宫内膜癌患者(内膜癌组),应用免疫组织化学法分别检测48例子宫内膜癌组织及15例正常子宫内膜(正常内膜组)组织中HIF-1α和EGF的表达情况,并用CD34抗体标记血管内皮细胞,测定微血管密度(MVD)值.对子宫内膜癌组织中HIF-1α、VEGF表达与临床参数及MVD进行相关性分析.结果 内膜癌组患者组织中HIF-1α和VEGF高表达率分别为54.17%、75.00%,均明显高于正常内膜组的6.67%、13.33%,组间比较差异均有显著统计学意义(P<0.01);内膜癌组与正常内膜组组织的MVD计数分别为(35.51±21.36)条、(3.66±4.52)条,组间比较差异有显著统计学意义(P<0.01);内膜癌组织中HIF-1α和VEGF表达与患者的年龄、临床分期、肌层浸润程度无关(P>0.05),与组织分化程度相关(P<0.05),而MVD与患者年龄、组织分化程度、临床分期、肌层浸润程度均有相关性(P<0.05).HIF-1α与VEGF的表达呈正相关(r=0.485,P=0.021);HIF-1α与MVD呈正相关(r=0.680,P=0.045);VEGF和MVD亦呈正相关(r=0.423,P=0.035).结论 子宫内膜癌组织中HIF-1α和VEGF均有高水平表达,并与肿瘤血管生成密切相关,两者可作为子宫内膜癌的肿瘤标志物,为临床抗肿瘤治疗提供新的方向.     

胰腺癌组织的平均微血管密度和血管内皮细胞生长因子的表达

目的:通过检测胰腺癌、慢性胰腺炎及正常胰腺标本中的平均微血管密度(MVD)及血管内皮细胞生长因子(VEGF)表达水平,了解胰腺癌肿瘤血管生成情况.方法:胰腺癌标本26例,慢性胰腺炎标本7例,癌旁正常胰腺组织26例(采用胰腺癌标本中无肿瘤浸润的组织),所有标本均行免疫组化染色.MVD检测为在200倍显微镜下计数5个视野最高数目的微血管,以其平均数表示.VEGF的定量标准为VEGF染色强度与阳性细胞百分比分值的乘积.统计分析采用t检验和二元变量相关分析法.结果:VEGF在胰腺癌、慢性胰腺炎及癌旁正常胰腺组织中均有表达.胰腺癌组织的MVD值及VEGF表达水平均明显高于慢性胰腺炎及癌旁正常胰腺组织(P<0.01).26例胰腺癌组织MVD值及VEGF表达水平在统计学上有相关性(Spearman法r=0.862,P<0.000;Pearson法r=0.855, P<0.000).结论:胰腺癌的微血管生成水平高于慢性胰腺炎及癌旁正常胰腺组织;胰腺癌组织内MVD值及VEGF表达水平在统计学上有相关性.     

HIF-1α，VEGF的表达及与乳腺癌生物学行为的关系

目的探讨乳腺癌组织中缺氧诱导因子-1α（hypoxia—inducible factor-1 alpha,HIF-1α）和血管内皮生长因子（vascular endothelial growth factor,VEGF）的表达及其与乳腺癌生物学行为之间的关系。方法采用免疫组化SP法检测乳腺纤维腺瘤、癌旁正常乳腺组织和乳腺癌中HIF-1α,VEGF蛋白的表达。结果HIF-1α蛋白在乳腺纤维腺瘤、癌旁正常乳腺组织中几乎无表达,乳腺导管原位癌中阳性率46．67％（14／30）,导管癌早期浸润中63．33％（19／30）,浸润性乳腺癌中76．67％（23／30）;乳腺癌中VEGF阳性率67．78％（61／90）;乳腺癌中HIF-1α表达与VEGF（r=0．64,P〈0．01）呈正相关,与c—erbB-2正相关（r=0．276,P〈0．01）;HIF-1α及其靶基因VEGF在乳腺癌组织中明显上调,HIF—1α表达与淋巴结转移、雌激素受体状态、组织学分级及临床分期相关。结论HIF-1α和VEGF的表达与乳腺癌的生物学行为有关,HIF—1α可能成为早期诊断乳腺癌、预测其转移的重要指标。     

P53及缺氧诱导因子-1α表达与结直肠癌血管生成的关系

目的探讨抑癌基因p53及缺氧诱导因子-1α(HIF-1α)的表达与结直肠癌血管生成及生物学行为的关系.方法采用免疫组织化学方法检测61例结直肠癌组织中p53、HIF-1α和血管内皮生长因子(VEGF)表达及微血管密度(MVD).结果结直肠癌组织中,HIF-1α阳性表达率为65.6%,其中弱、中、强阳性表达率分别为27.9%、19.7%、18.0%,p53和VEGF阳性表达率分别为42.6%和49.2%,MVD平均为28.7±12.9.HIF-1α、VEGF表达及MVD与肿瘤浸润深度、淋巴结转移、肝转移和Dukes分期密切相关.p53表达与淋巴结转移和肝转移密切相关.p53、VEGF阳性表达率及MVD均与HIF-1α阳性表达程度成正相关(相关系数分别为0.261,P＜0.05;0.591,P＜0.01;0.795,P＜ 0.01).结论HIF-1α/VEGF通路在结直肠癌组织血管 生成过程中起重要作用,p53基因失活可能经HIF-1α/VEGF通路促进肿瘤血管生成.p53基因失活、HIF-1 α、VEGF过表达及高MVD与结直肠癌的不良生物学行为有关.     

血管内皮生长因子与脑膜瘤瘤周水肿的关系

目的　探讨脑膜瘤血管内皮生长因子 (VEGF)表达与肿瘤血管生成和瘤周脑水肿的关系。方法　采用免疫组织化学技术检测 40例脑膜瘤VEGF表达和微血管密度 ,用双盲法对染色结果及术前MRI和脑血管造影资料进行评估。结果　VEGF表达强阳性组与表达阴性组相比 ,水肿指数(EI =4 6与EI=1 5 ,P =0 0 0 3) ,脑水肿发生率 (88 2 %与 41 7% ,P =0 0 2 3 68)、微血管密度 (MVD =53 0与MVD =2 6 5 ,P =0 0 1 8)的差异均有显著意义 ;有软膜供血组与无软膜供血组相比水肿指数差异有显著意义 (EI=4 4与EI=1 8,P =0 0 4 4 ) ;肿瘤与脑组织重度粘连组较轻度粘连的脑水肿发生率差异有显著意义 (88 9%与 45 5 % ,P =0 0 0 4 1 7) ;肿瘤大小与水肿指数无相关关系 (r =0 2 64 ,P >0 0 5)。结论　脑膜瘤的VEGF表达、肿瘤血管生成及瘤周脑水肿形成之间有重要关系 ;VEGF能促进脑膜瘤的肿瘤血管生成及瘤周脑水肿的形成 ;肿瘤有软膜供血的、与脑组织粘连严重的脑水肿发生率高 ;肿瘤大小与脑水肿程度无相关关系     

血管内皮生长因子在原发性食管鳞状细胞癌中的表达及意义

目的：探讨原发性食管鳞状细胞癌中血管内皮生长因子（VEGF）的表达情况及其与肿瘤血管生成和预后的关系。方法：采用免疫组织化学染色方法,分别以抗VEGF多克隆抗体的抗F－VIII单克隆抗体检测82例手术切除标本中VEGF的表达水平和肿瘤微血管密度（MVD）值,分析二者之间及与患者预后的相关性。结果：全组病例中,肿瘤细胞VEGF阳性率为63．4％（52／82）VEGF阴性和VEGF阳性的3年和5年生存率分别为53．4％和46．6％;19．2％和11．5％。MVD中位值为37个/mm^2(9-150个／mm^2)。VEGF的表达水平与MVD值密切相关（P＝0．001）。MVD与患者的预后显著相关（P＝0．017）。随着VEGF表达水平增高,患者预后明显恶化（P＝0．000）。结论：VEGF在食管鳞状细胞癌的肿瘤血管生成过程中发挥重要的调控作用。多因素预后分析表明VEGF的表达可作为食管鳞状细胞癌患者独立的预后判断指标。     

缺氧诱导因子-1α、血管内皮生长因子在婴幼儿血管瘤的表达及意义

目的:探讨缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)在婴幼儿血管瘤和婴幼儿正常皮肤的表达情况。方法:采用免疫组织化学S-P染色法,检测HIF-1α、VEGF在婴幼儿血管瘤及正常皮肤的表达。结果:HIF-1α、VEGF表达水平在血管瘤与正常皮肤之间差异有统计学意义(P<0.05)。血管瘤中HIF-1α与VEGF表达呈正相关。结论:HIF-1α与VEGF在血管瘤中呈阳性表达,可能参与促进血管瘤血管生成。     

缺氧诱导因子-1α在浸润性宫颈鳞癌组织中的表达及其与微血管密度的关系

目的:研究缺氧诱导因子1α(HIF-1α)在浸润性宫颈鳞癌组织中的表达及其与微血管密度的关系。方法:采用免疫组化EliVision法检测HIF-1α在20例正常宫颈和52例浸润性宫颈鳞癌组织中的表达特点,CD34抗体标记血管内皮细胞并计数MVD。结果:HIF-1α在浸润性癌组织中的阳性表达率为53.84%,明显高于正常组织表达率25.00%(P<0.05);FIGOⅡ期和淋巴结转移肿瘤组织HIF-1α表达明显高于Ⅰ期组和淋巴结未转移组(P<0.05)。肿瘤组织的MVD计数为38.85±4.04,明显高于正常对照11.95±2.31 (P< 0.05);Ⅱ期肿瘤MVD计数明显高于Ⅰ期,淋巴结转移组MVD计数明显高于未转移组(P< 0.05)。肿瘤组织中MVD计数与HIF-1α的表达水平正相关(r=0.31,P=0.02)。结论:浸润性宫颈鳞癌组织中HIF-1α表达与血管生成密切相关。HIF-1α过表达和高MVD计数与浸润性宫颈鳞癌的恶性生物学行为有关,可成为预测浸润性宫颈鳞癌侵袭转移的重要指标。     

水通道蛋白 1、血管内皮生长因子及微血管密度 在胃癌组织中的表达及临床意义

目的??探讨水通道蛋白 1（AQP1） 、血管内皮生长因子（VEGF） 、微血管密度（MVD）在胃癌组织中 的表达及 AQP1、VEGF、MVD 之间的相关性和临床意义。方法??选取郑州大学第二附属医院胃癌标本 79 例。每 例分别取癌组织和癌旁组织（距肿瘤边缘 <2?cm 处） ,采用免疫组织化学方法检测 AQP1、VEGF 与 MVD 在人 胃癌组织和癌旁组织中的表达,对 AQP1、VEGF 与 MVD 的表达及其与临床病理参数的关系进行分析。结果? AQP1、VEGF 在胃癌组织与癌旁组织比较,差异有统计学意义（ P ?<0.05） ,胃癌组织高于癌旁组织 ; MVD 在 胃癌组织的表达与癌旁组织比较,差异有统计学意义（ P ?<0.05） ,胃癌组织高于癌旁组织 ; AQP1、VEGF 及 MVD 与胃癌的分化、淋巴结转移、脉管癌栓及分期相关（ P ?<0.05） ; 胃癌组织中 AQP1 与 VEGF 的表达呈正 相关（ r =0.497, P ?<0.05） ; 在 VEGF 阳性表达的胃癌组织中, MVD 表达高于 VEGF 阴性表达的组织（ P ?<0.05） ,在 AQP1 阳性表达的胃癌组织中,MVD 表达高于 AQP1 阴性表达的组织（ P ?<0.05） 。结论??AQP1、VEGF、MVD 存在协同作用, 与肿瘤的发生、发展密切相关。