缺氧诱导因子-1α在腹主动脉瘤中的表达及其意义

目的　观察缺氧诱导因子 (HIF) 1α信号传导途径在腹主动脉瘤 (AAA)中的表达 ,探讨AAA的发病机制。方法　选取AAA标本 2 2例 ,以 5例正常腹主动脉 (NA )标本作对照。采用Northern杂交、Western蛋白印迹及免疫组织化学方法检测HIF 1α的mRNA及蛋白产物表达 ,Western蛋白印迹和免疫组织化学方法检测血管内皮生长因子 (VEGF)的表达 ,免疫组织化学抗CD3 4染色法检测微血管密度 (MVD)。结果　AAA组织中HIF 1α的mRNA及蛋白产物表达明显高于NA(P <0 .0 1) ;VEGF表达亦明显增强 (P <0 .0 1) ,与HIF 1α表达呈显著正相关 (r值为0 .783 ,P <0 .0 1)。HIF 1α表达主要分布在AAA中层血管平滑肌细胞 (VSMC)及外膜处 ,与VEGF的分布部位基本一致。AAA中微血管密度明显增加 (P <0 .0 1) )。结论　HIF 1α在AAA发病过程中可能发挥重要作用 ,其作用机制可能是通过促进VEGF的表达而实现的。     

缺氧诱导因子-1α及相关基因在破裂性腹主动脉瘤中的表达及意义

目的研究缺氧诱导因子(hypoxia-inducible factor-1α,HIF-1α)信号转导通路在破裂性腹主动脉瘤(ruptured abdominal aorta aneurysm,RAAA)中的表达,探讨AAA破裂的分子基础。方法选取RAAA标本18例,以20例AAA标本作对照。采用Northern杂交、Western蛋白印迹及免疫组织化学方法检测HIF-1α的mRNA及蛋白产物表达,Western蛋白印迹和免疫组织化学方法检测血管内皮生长因子(VEGF)的表达,免疫组织化学抗CD34染色法检测微血管密度(microvessel density, MVD)。结果 RAAA组织中HIF-1α mRNA表达明显高于AAA组(P<0．01),免疫组化染色表明 HIF-1α阳性细胞百分率为(40．9±10．8)％,与AAA(17．6±5．2)％比较差异有统计学意义(P< 0．01);VEGF表达亦明显增强(P<0．01),与HIF-1α表达呈显著正相关(r值为0．725,P<0．01)。 HIF-1α阳性表达主要分布在AAA中层平滑肌细胞及外膜。RAAA中微血管密度明显增多(P< 0．01)。结论 HIF-1α及其相关基因过表达在RAAA的分子机制中可能发挥重要作用。     

大肠腺癌中辅助蛋白同源的磷脂酶基因和缺氧诱导因子-1α的表达及与血管内皮生长因子的关系

目的研究大肠腺瘤和腺癌组织中与张力蛋白和辅助蛋白同源的磷脂酶mRNA(PTENmRNA)、缺氧诱导因子-1αmRNA(HIF-1αmRNA)、血管内皮生长因子(VEGF)蛋白的表达及其临床意义。方法PTENmRNA、HIF-1αmRNA检测采用原位杂交技术,应用免疫组织化学方法检测VEGF蛋白的表达。结果HIF-1αmRNA阳性表达为棕黄色杂交颗粒,主要定位于肿瘤组织坏死边缘区域的肿瘤细胞胞浆中。62例大肠腺癌标本中,PTENmRNA、HIF-1mRNA、VEGF蛋白阳性表达率分别为51.6%、67.7%、59.7%。18例大肠腺瘤标本中,PTENmRNA、HIF-1mR-NA、VEGF蛋白阳性表达率分别为77.8%、44.4%、33.3%。大肠腺癌VEGF蛋白阳性表达率显著高于大肠腺瘤(P<0.05),而PTENmRNA则相反。有局部复发、淋巴结转移、肝转移、Dukes分期高的大肠腺癌组织中HIF-1mRNA阳性表达率显著增高,而PTENmRNA表达则相反。HIF-1α与VEGF呈显著正相关(χ2=4.751,P<0.05);HIF-1αmRNA表达与PTENmRNA呈显著负相关(χ2=21.84,P<0.01);HIF-1αmRNA与VEGF无相关性(χ2=2.597,P>0.05)。结论抑癌基因PTEN的低表达或突变缺失诱导HIF-1α及其靶基因VEGF的过度表达在大肠腺瘤-腺癌发展过程中起重要作用。     

Rac1、缺氧诱导因子-1α、血管内皮生长因子在胃癌组织中的表达和血管生成的关系

我们采用免疫组织化学方法检测60例胃癌组织和30例正常胃黏膜组织中Rac1、缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)的表达,并用CD34标记血管内皮细胞,计算微血管密度(MVD),探讨Rac1、HIF-1α、VEGF与肿瘤血管生成的关系.     

低氧诱导因子-1α和p53在肝脏低氧损伤中的作用

目的:探讨低氧诱导因子-1α(HIF-1α)、p53及细胞凋亡在大鼠肝脏低氧损伤中的作用。方法:雄性SD大鼠64只,随机分为对照组和肝动脉结扎组(n=32)。各组均行剖腹、肝脏骨骼化及肝素林格液肝脏灌注,肝动脉结扎组行肝动脉结扎术。术后1、3、7、14d留取肝脏组织及血标本。采用反转录-聚合酶链反应(RT-PCR)法检测肝脏组织HIF-1αmRNA表达水平;免疫组织化学方法检测肝脏组织HIF-1α及p53蛋白表达水平;TUNEL法检测肝细胞凋亡。自动生化分析仪检测血清丙氨酸转氨酶(ALT)活性以评价肝损伤。结果:肝动脉结扎后肝组织HIF-1αmRNA及HIF-1α蛋白较对照组明显增高,分别于结扎后3d和7d达峰值[0.834±0.129比0.372±0.048,(7.8±3.1)%比(2.1±1.7)%,P均<0.01]。肝动脉结扎组肝细胞p53均明显高于对照组(P<0.05或P<0.01),于术后7d达峰值[(41.2±9.1)%比(5.2±4.3)%,P<0.01]。肝动脉结扎组肝细胞凋亡指数均明显高于对照组(P<0.05或P<0.01),于术后7d达峰值[(21.3±7.4)%比(3.7±3.5)%,P<0.01]。肝动脉结扎组ALT水平均明显高于对照组。HIF-1α、p53蛋白表达及凋亡细胞都主要位于肝小叶中央静脉周围。结论:HIF-1αmRNA表达增加、HIF-1α蛋白积聚以及p53促凋亡途径的激活在肝脏低氧损伤中可能发挥重要作用。     

外泌体微小RNA-let-7c靶向结合AGO1调控多发性骨髓瘤血管生成

目的探究多发性骨髓瘤外泌体中微小RNA(miR)-let-7c对血管内皮细胞生物学功能的影响及机制。方法从多发性骨髓瘤初治患者的骨髓活检标本中原代培养出骨髓瘤内皮细胞, 慢病毒感染后建立miR-let-7c过表达或抑制内皮细胞稳染株, 使用细胞计数试剂盒(CCK-8)、Transwell迁移实验及小管形成实验检测感染后内皮细胞的生物学行为改变, 并采用生物学信息技术预测miR-let-7c与argonaute1(AGO1)蛋白的靶向调控区, 双荧光素酶报告基因实验论证两者之间的靶向调控作用。构建多发性骨髓瘤移植瘤小鼠模型, 采用实时定量聚合酶链式反应(qRT-PCR)检测各组移植瘤组织中miR-let-7c的表达水平, 免疫组织化学标记内皮细胞表面CD31来计算各组肿瘤微血管密度(MVD);采用蛋白印迹法检测各组移植瘤组织中缺氧诱导因子1α (HIF-1α)、AGO1及血管内皮生长因子(VEGF)的蛋白表达;采用qRT-PCR检测各组移植瘤组织中HIF-1α、AGO1及VEGF的mRNA表达。两组均数间用独立样本t检验, 两组以上均数的比较用方差分析。结果用CCK-8试剂盒检测发现, ...     

缺氧诱导因子-1α和缺氧诱导因子-2α在人非小细胞肺癌表达的比较

目的 观察缺氧诱导因子(HIF)-1α和HIF-2α在人非小细胞肺癌(NSCLC)中的表达并探讨其临床意义.方法 采用实时定量聚合酶链反应(PCR)和Western blot法检测不同缺氧时程HIF-1α和HIF-2α的mRNA和蛋白表达,比较两者差异.进一步运用免疫组织化学技术检测两者在140例NSCLC患者标本中的表达,结合相关临床病理因素探讨其临床意义.结果 在缺氧条件下,HIF-1α蛋白在4h即迅速升至峰值,之后快速下降,而HIF-2α蛋白则表现为逐步上调并持续维持在较高表达水平.在140例NSCLC的标本研究中,HIF-2α与肿瘤大小、淋巴结转移和TNM分期关系密切(P<0.05),并且是预后的不良因素(P<0.05).结论 HIF-2α可能是肿瘤适应乏氧环境的主要调节因子,相较于HIF-1α、HIF-2α在NSCLC的靶向治疗研究中更具有价值.     

反义HIF-1α基因治疗人肝癌裸鼠移植瘤的实验研究

摘要：目的:探讨反义HIF-1α基因对人肝癌裸鼠皮下移植瘤的影响及其相关机制。方法:使用人原发性肝癌细胞株SMMC-7721建立人肝癌裸鼠皮下移植瘤动物模型。待肿瘤生长至直径约0.4cm时，将荷瘤裸鼠随机分为3组，分别注射生理盐水、质粒PcDNA3和HIF-1α/PcDNA3B，质粒用脂质体DOTAP介导转染细胞。观察各组动物的肿瘤生长曲线；取肿瘤标本作免疫组织化学检查（SABC法）及蛋白质印迹检查，检测各组肿瘤的VEGF和HIF-1α表达及微血管密度（MVD）和细胞凋亡。结果:HIF-1α/PcDNA3B治疗组各时点的肿瘤体积，以及肿瘤组织中HIF-1α蛋白、MVD，VEGF表达均低于对照组，而细胞凋亡指数高于对照组，差异均有显著性（P＜0.05）。结论:通过阻断癌细胞的缺氧适应途径，反义HIF-1α基因治疗肝癌有抑制肿瘤生长、抑制肿瘤血管生成及诱导细胞凋亡的作用。     

缺氧诱导因子-1α、血管内皮生长因子在兔早期肝硬化形成过程中的表达

目的 观察兔早期肝硬化形成过程中缺氧诱导因子(HIF)-α、血管内皮生长因子(VEGF)在肝组织的表达.方法 实验设四氯化碳诱导和正常对照组各15只新西兰大白兔,分别每2周处死实验组3只,正常对照组3只,同时进行常规病理学检测及免疫组织化学检测.结果 HIF-α免疫组织化学染色阳性细胞主要位于肝细胞的细胞质中,部分细胞核中也有表达.VEGF免疫组织化学染色阳性细胞位于肝细胞的细胞质、细胞核和血管内皮细胞中.HIF-1α、VEGF阳性表达均呈棕黄色颗粒.HIF-1α、VEGF在早期肝硬化期(12周末)阳性表达数均为10例;在肝纤维化期(8周末)阳性表达数均为9例;在肝炎期(4周末)阳性表达数均为4例.HIF-1α mRNA、VEGF mRNA在3个时期表达水平差异有统计学意义(P＜0.05),并随时间延长其阳性表达率增加.结论 通过对兔早期肝硬化形成过程中HIF-1α、VEGF在肝组织的表达的观察,为临床基因靶点治疗肝脏疾病提供实验基础.     

Ang2,HIF-1α及VEGF对肝癌血管形成的影响

摘要：目的:探讨促血管生成素2（Ang2）、缺氧诱导因子1α（HIF-1α）及血管内皮生长因子（VEGF）与肝细胞癌血管形成的关系。方法:检测52例肝癌组织中Ang2，HIF-1α及VEGF mRNA及蛋白的表达，对共表达的肝癌组织进行微血管计数。结果:RT-PCR 显示,52例肝癌组织中有38例共表达Ang2mRNA，HIF-1αmRNA 和VEGF mRNA，且两两之间呈明显正相关(分别为r=0.783，P<0.01；r=0.427，P<0.05；r=0.433，P<0.05)；免疫组化发现,52例肝癌组织36例共表达Ang2，HIF-1α和VEGF蛋白。共表达Ang2 mRNA，HIF-αmRNA 和VEGF蛋白的38例肝癌组织中,平均微血管数［(45.4±8.90) 个/HP］,明显高于非共表达组［(13.6±3.30)个/HP］（P<0.05)。结论:Ang2，HIF-1α和VEGF与肝癌的新生血管形成有关；肿瘤组织缺氧可能是其始动因素。     

胃癌组织缺氧诱导因子-1α与胰岛素样生长因子-Ⅱ表达对肿瘤血管生成的影响及预后评估

目的探讨胃癌组织中缺氧诱导因子-1α（HIF-1α）及胰岛素样生长因子-Ⅱ（IGF—Ⅱ）mRNA的表达水平及其与血管生成和预后的关系。方法采用原位杂交检测118例胃癌组织中HIF-1αmRNA和IGF—ⅡmRNA的表达情况,用免疫组化SP法标记CD34单克隆抗体,计算肿瘤微血管密度（MVD）。结果HIF—1αmRNA和IGF—ⅡmRNA在胃癌组织中的阳性表达率分别为49．2％和47．4％。且与胃癌临床侵袭转移病理指标均相关。HIF-1αmRNA和IGF-ⅡmRNA阳性表达者的MVD值,均高于阴性表达者;HIF-1αmRNA和IGF-ⅡmRNA表达呈正相关,MVD分别与HIF-1αmRNA和IGF—ⅡmRNA的表达水平呈正相关。HIF—1αmRNA和IGF—ⅡmRNA表达阳性及MVD值≥41．5个／0．72mm^2的患者的平均生存时间及5年生存率均低于表达阴性及MVD值〈41．5个／0．72mm^2者。结论HIF-1α与IGF—Ⅱ对胃癌的浸润转移,特别是对肿瘤血管形成具有重要作用,可作为预测患者预后和指导治疗的新途径。     

血管瘤中缺氧诱导因子-1α的表达和血管生成的研究

目的　探讨缺氧诱导因子- 1α(HIF-1α)在血管瘤中的表达以及其和血管内皮细胞生长因子 (VEGF)、新生微血管密度 (MVD)的关系。方法　采用免疫组化SP法检测 2 8例婴幼儿血管瘤中HIF 1α、VEGF的蛋白表达和MVD。结果　 2 8例血管瘤中HIF-1α、VEGF的蛋白表达阳性率分别是6 4 % ,71.4 %。其中增生期和消退期HIF 1α阳性率分别为 87.5 %、33.3% (P <0.0 1) ,VEGF阳性率分别为 93.7%、4 1.7% (P <0.0 1) ;MVD分别为 73 4± 14 6 3、30 2± 9 1(P <0.0 1)。HIF 1α蛋白表达与VEGF成正相关 (P <0.0 1) ,HIF 1α和VEGF与MVD都成正相关 (P <0.0 1)。结论　血管瘤增生期血管内皮细胞存在着特殊的缺氧微环境 ,并通过HIF-1α表达水平提高调节VEGF等血管生成相关因子表达水平升高 ,促进了新生微血管生成     

Hypoxia-inducible factor 1 alpha expression correlates with angiogenesis and unfavorable prognosis in bladder cancer

INTRODUCTION AND OBJECTIVES: Hypoxia-inducible factor 1 alpha (HIF-1 alpha) is a critical regulatory protein of cellular response to hypoxia and is closely related to the triggering of the angiogenic process. We examined the relationship between hypoxia and angiogenesis, as well as their prognostic impact in patients with urothelial bladder cancer. METHODS: The immunohistochemical expression of HIF-1 alpha was evaluated in 93 formalin-fixed paraffin-embedded primary transitional cell carcinoma tissue samples. HIF-1 alpha was recognized through nuclear staining of positive cells. The angiogenic profile was individually assessed immunohistochemically using a monoclonal antibody to vascular endothelial growth factor (VEGF) and microvessel density (MVD) was calculated with immunohistochemical staining of the adhesion molecule CD31 of the endothelial cells. RESULTS: A significant positive association between HIF-1 alpha immunoreactivity and histological grade (p=0.009) was found. VEGF and MVD were closely related to tumor grade (p=0.06 and p<0.001) and clinical stage (p=0.04 and p<0.01, respectively). HIF-1 alpha was significantly correlated with VEGF expression (p=0.01) and MVD (p<0.001). Patients characterized by HIF-1 alpha overexpression had significantly worse overall (p=0.009) and disease-free survival (p=0.03). When HIF-1 alpha, histologic grade and stage were included in multivariate Cox regression analysis, HIF-1 alpha emerged as an independent prognostic factor (p=0.02) along with grade and stage, but lost its independent prognostic value after the inclusion of angiogenic factors in the multivariate model. In the subgroup of patients with T1 disease, HIF-1 alpha emerged as a significant negative predictor of the time to first recurrence. CONCLUSIONS: HIF-1 alpha and angiogenesis markers may play an important predictive and prognostic role in patients with bladder cancer. HIF-1 alpha may be of biologic and clinical value as its overexpression is related to up-regulation of VEGF, the stimulation of angiogenesis and worse prognosis.     

缺氧诱导因子-1α在大鼠门静脉高压胃病中的作用研究

摘要：目的:通过检测缺氧诱导因子-1α(HIF-1α)在门静脉高压胃病（PHG）大鼠胃壁中的表达，探讨其对PHG病变所起的作用。方法:通过门静脉部分缩窄方法制备PHG大鼠模型，并设立假手术组（SO）作为对照，免疫组化法检测HIF-1α和血管内皮生长因子(VEGF)及CD34在大鼠胃壁组织中的表达，并对CD34阳性血管进行微血管密度 (MVD) 计数。结果:HIF-1α，VEGF和CD34在PHG组大鼠胃壁中的表达均明显高于SO组（P<0.01）。结论:HIF-1α的过度表达可能在PHG发病机制中起重要作用。     

低氧诱导因子1α及血管内皮生长因子在前列腺癌中的表达及意义

目的:观察低氧诱导因子1α(H IF-1α)及血管内皮生长因子(VEGF)在前列腺癌(PCa)中的表达及意义。方法:32例PCa患者,根据G leason评分,将≥7分者设为高G leason评分组(n=12),<7分者为低G leason评分组(n=20)。良性前列腺增生(BPH)16例,BPH伴高级别前列腺上皮内瘤(PIN)15例,正常前列腺组织(NP)12例。采用免疫组化染色CD34观察各组组织中微血管密度(MVD)及H IF-1α、VEGF的表达情况。结果:PCa、PIN中H IF-1α阳性表达率分别为62.5%、60.0%,较BPH(6.3%)及NP(0)高,差异有统计学意义(P<0.05)。PCa、PIN中VEGF阳性表达率分别为78.1%、73.3%,较BPH(18.7%)及NP(8.3%)高,差异亦有统计学意义(P<0.05)。PCa的MVD为66.9±18.0,明显高于BPH(28.3±6.9)及NP(15.3±2.9)(P<0.05)。高G leason评分组H IF-1α、VEGF阳性率及MVD值均高于低G leason评分组,差异均有显著性(P<0.05)。结论:H IF-1α及VEGF过表达是PCa形成的早期事件,与PCa密切相关。     

Experimental varicocele induces hypoxia inducible factor-1alpha, vascular endothelial growth factor expression and angiogenesis in the rat testis

PURPOSE: In this study we investigated hypoxia inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) expression, and angiogenesis in an experimental model of varicocele in the rat testis. MATERIALS AND METHODS: A total of 30 adult male Sprague-Dawley rats were investigated in 3 groups, namely varicocele group 1 (13), sham operated group 2 (9) and control group 3 (8). At 30 days after surgery was completed in groups 1 and 2 orchiectomy was performed in all rats. Histological findings in the left testicles of rats from each group were compared. HIF-1alpha and VEGF expression was immunohistochemically studied and CD31 panendothelial antigen was used to identify the number of microvessels, that is microvessel density (MVD), in paraffin embedded sections of testis tissue. Data were analyzed using the chi-square test, Fisher's exact test, 1-way ANOVA and the Tukey HSD test for post hoc comparison. RESULTS: HIF-1alpha expression was detected in 12 specimens (92.3%) in group 1, 4 (44.4%) in group 2 and 2 (25%) in group 3. The frequency of HIF-1alpha positivity in group 1 was significantly higher than the rates in groups 2 (p = 0.023) and 3 (p = 0.003). VEGF expression was detected in 8 specimens (61.5%) in group 1 but none of the group 2 or 3 specimens were VEGF positive. The frequency of VEGF positivity in group 1 was significantly higher than that in groups 2 (p = 0.006) and 3 (p = 0.007). Mean MVD +/- SD in group 1 was 7.53 +/- 1.50 (range 6 to 12), and findings in groups 2 and 3 were 5.88+/-1.45 (range 4 to 8) and 5.12 +/-1.12 (range 4 to 7), respectively. Mean MVD in group 2 was higher than in group 3 but this difference was not significant (p = 0.509). Mean MVD in group 1 was significantly higher than the mean values in groups 2 (p = 0.030) and 3 (p = 0.002). CONCLUSIONS: Previous study of experimental varicocele models in rats documented HIF-1alpha and VEGF expression combined with angiogenesis in the testis. The results of this study show that varicocele can lead to tissue hypoxia and related pathophysiological events, such as angiogenesis.     

Expression of hypoxia-inducible angiogenic proteins (hypoxia-inducible factor-1alpha, vascular endothelial growth factor, and E26 transformation-specific-1) and plaque hemorrhage in human carotid atherosclerosis

OBJECT: Plaque hemorrhage in carotid atherosclerosis promotes plaque progression, resulting in cerebrovascular disease. Hypoxia inducible factor-1alpha (HIF-1alpha) induces angiogenesis via the expression of vascular endothelial growth factor (VEGF) and E26 transformation-specific-1 (Ets-1). The authors investigated human carotid plaques to determine whether these hypoxia-inducible angiogenic proteins play a major role in intraplaque angiogenesis and hemorrhage. METHODS: The expression of HIF-1alpha, VEGF, and Ets-1 was analyzed using immunohistochemistry and Western blotting in 29 human carotid plaques obtained at carotid endarterectomy. The authors investigated the relationship between plaque characteristics and clinical symptoms. RESULTS: A higher incidence of plaque hemorrhage was observed in plaques associated with symptoms than in those without symptoms (p = 0.03). Hypoxia-inducible factor-1alpha, VEGF, and Ets-1 coexisted in the deep layer of plaque, where angiogenesis was remarkably developed; the expression levels of HIF-1alpha, VEGF, and Ets-1 were significantly enhanced in the main lesion of the plaque (p < 0.01). Symptomatic plaques showed higher expression of VEGF (p = 0.04) than asymptomatic plaques. Plaques with hemorrhage showed a higher incidence of plaque ulcer (p = 0.001) and higher expression of Ets-1 (p = 0.03) than those without hemorrhage. Moreover, significantly increased expressions of VEGF (p = 0.01) and Ets-1 (p = 0.006) were observed in plaques with not only hemorrhages but also ulcers and severe stenosis. CONCLUSIONS: The findings in this study suggest that hypoxia-inducible angiogenic proteins in human carotid atherosclerosis promote intraplaque angiogenesis, which can induce plaque hemorrhage and progression.