血管新生在急慢性白血病中的研究及临床评价

 目的 观察急慢性白血病患者骨髓血管新生情况,分析血管内皮细胞生长因子（VEGF）及其受体（VEGFR）在急慢性白血病患者中的表达及意义。方法 应用vWF标记免疫组化法观察骨髓血管新生情况,ELISA法检测血浆VEGF水平,流式细胞术（FACS）分析新鲜分离的白血病细胞表面VEG-FR表达。结果 32例初诊白血病患者骨髓微血管密度（MVD）较对照组明显增高,治疗后获缓解者骨髓MVD较治疗前明显降低,与对照组相比差异无统计学意义。32例初诊白血病患者血浆VEGF含量明显高于对照组,化疗后获完全缓解（CR）的急性白血病患者血浆VEGF含量降低至正常水平。31例白血病患者骨髓单个核细胞（BMMNC）27例不同程度表达VEGFR,而5例正常对照骨髓均无VEGFR表达。结论 初诊白血病患者存在骨髓血管新生及血浆VEGF水平增高,白血病细胞不同程度表达VEGFR,阻断VEGF-VEGFR信号通路可成为白血病治疗新的靶点。     

白血病患者骨髓血管新生的研究

目的研究白血病患者的骨髓血管新生程度和骨髓液血管内皮生长因子(VEGF)水平的相关性。方法采用EnVison免疫组化法,用兔抗人vWF(vonWille蛳brandfactor)多克隆抗体标记骨髓内皮细胞;采用ELISA法检测骨髓液中VEGF的含量。结果骨髓MVD在各实验组均明显高于正常对照组(P<0.05)。骨髓液VEGF水平在AML、CML组明显高于正常对照组(P<0.01),ALL组与正常对照组比较差异无显著性(P>0.05)。对各白血病组的MVD和VEGF含量进行了相关性分析,结果表明在AML或CML组中VEGF水平和MVD呈正相关性(分别为r=0.648,P<0.05;r=0.733,P<0.05)。结论白血病患者骨髓微血管数增多,提示血管新生在白血病发病中有重要作用,VEGF可能在一定程度上对血管新生有促进作用。     

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。     

Argonaute 2与多发性骨髓瘤血管生成临床关系研究

目的 了解Argonaute 2(Ago2)在多发性骨髓瘤(MM)骨髓组织中的表达水平,探讨其与骨髓内血管生成的关系.方法 应用改良的甲基丙烯酸甲酯(MMA)单体塑料包埋法对59例MM患者及16例正常对照进行骨髓活组织检查的组织制片,采用EnVision免疫组织化学二步法检测MM患者及正常对照骨髓组织中Ago2蛋白表达水平及微血管密度(MVD),并用Western blot方法检测8例MM患者和3例正常对照Ago2蛋白表达水平.结果 Western blot检测显示,Ago2蛋白在MM患者骨髓组织中表达高于正常对照(1.35±0.19比0.15±0.03,t=-19.883,P＜0.001).免疫组织化学结果表明,59例MM患者骨髓组织中Ago2蛋白阴性和阳性分别为9例和50例,对照组16例均为阴性,两组差异有统计学意义(x2=42.586,P＜ 0.001).MM患者中,Ago2阳性组β2-微球蛋白高于阴性组,差异有统计学意义(Z=-2.014,P=0.042).MM患者骨髓组织中MVD为7.89±4.88,正常对照组为2.16±1.32,两组之间差异有统计学意义(t=4.63,P＜0.001).MM患者骨髓组织中Ago2蛋白表达水平与MVD具有相关性(r=0.461,P=0.023).Ago2蛋白阳性组MVD较阴性组增高,差异有统计学意义(t=2.71,P=0.009).结论 Ago2在MM患者骨髓组织中表达水平明显升高,可能参与了MM患者的病理发生,并对骨髓内异常新生血管生成起一定的调节作用.     

miR-455-3p在胃癌中的表达及其与MVD和VEGF表达的相关性研究

目的 探讨miR-455-3p在胃癌中的水平及其与微血管密度（MVD）和血管内皮生长因子（VEGF）表达的关系。方法 采用实时荧光定量PCR检测80例胃癌组织（胃癌组）中miR-455-3p水平,免疫组化法检测80例胃癌组织中MVD和VEGF表达,并分析胃癌组织中miR-455-3p水平与MVD和VEGF表达的相关性。选取同期的37例浅表性胃炎和12例正常胃黏膜组织作对照（对照组）。结果 胃癌组的miR-455-3p水平为1.16±0.59,低于对照组的2.61±0.88,差异有统计学意义（P＜0.05）;胃癌组的VEGF阳性表达率和MVD分别为71.3％（57／80）和54.9±7.3,均高于对照组的28.6％（14／49）和27.5±6.1,差异均有统计学意义（P＜0.05）;胃癌组织中miR-455-3p水平与VEGF表达（r=-0.783,P＜0.05）和MVD（r=-0.824,P＜0.05）均呈负相关。结论 miR-455-3p在胃癌中为低表达,并与VEGF表达和MVD呈负相关,有可能在调控胃癌组织血管生长中发挥重要作用。     

多发性骨髓瘤骨髓组织中p53与血管新生的关系

目的　研究多发性骨髓瘤骨髓组织中 p5 3基因和VEGF表达与血管新生的关系。 方法　应用原位杂交及免疫组化染色技术对 4 2例多发性骨髓瘤患者骨髓组织中VEGF、p5 3及微血管密度 (MVD)进行检测。结果　VEGF的阳性率为 5 2 .4 % (2 2 /4 2 ) ,p5 3的阳性率为2 1.4 % (9/4 2 )。经连续切片对比及统计学分析显示 ,p5 3的表达与VEGF的表达显著相关 (P <0 .0 5 ) ;并且p5 3阳性组MVD显著高于阴性组MVD(P <0 .0 1) ,VEGF阳性组MVD显著高于阴性组MVD(P <0 .0 1)。结论　多发性骨髓瘤骨髓组织中 p5 3突变可以上调VEGF的表达 ,促进血管新生。     

食管鳞癌p53、TSP-1、VEGF表达与肿瘤血管生成

目的 探讨食管鳞癌中p53、血小板反应蛋白-1（TSP-1）和血管内皮生长因子（VEGF）的表达与肿瘤血管生成的关系。方法 利用免疫组织化学（SP法）方法检测68例食管鳞癌组织中p53、TSP-1和VEGF的表达及肿瘤内微血管密度（MVD）计数。结果 食管鳞癌组织中p53、TSP-1、VEGF阳性率分别为72.06%、29.41%和64.71%。p53与TSP-1呈负相关（rs =－1.000,P〈0.01）,与VEGF呈正相关（rs =1.000,P〈0.01）。TSP-1阳性组MVD为18.37±4.86,TSP-1阴性组MVD为29.80±6.35（t = 2.735,P〈0.01）,TSP-1与MVD呈负相关（rs =－0.783,P〈0.01）。结论 p53通过调节TSP-1和VEGF的表达,促进了食管鳞癌组织新生血管的生成。     

外周T细胞淋巴瘤中VEGF表达和MVD水平及其临床意义

目的 探讨外周T细胞淋巴瘤（PTCL）中血管内皮细胞生长因子（VEGF）表达和微血管密度（MVD）水平及其与临床特征和预后的相关性。方法 采用免疫组化法检测47例初治PTCL患者肿瘤组织中VEGF及MVD表达,分析其与临床特征、疗效和预后的关系。结果 PTCL组织中VEGF阳性表达率为93.6％（44／47）,其中高表达率为55.3％（26／47）;MVD值为（97.99±48.45）／mm²,其中MVD高表达者24例。MVD水平与VEGF表达呈正相关（r=0.329,P=0.024）。VEGF表达与临床分期、骨髓浸润、国际预后指数 （IPI）评分及ECOG评分相关（P＜0.05）;MVD仅与临床分期相关（P=0.028）。VEGF高表达组的5年生存率、5年无进展生存率分别为11.5％、11.5％,低于VEGF低表达组的57.1％、28.6％,差异均有统计学意义（P＜0.05）。MVD高水平组的5年生存率、5年无进展生存率分别为20.8％、8.3％,低于MVD低水平组的43.5％、 30.4％,差异均有统计学意义（P＜0.05）。Log-rank检验显示,VEGF表达、MVD水平、临床分期、IPI评分、LDH表达水平、骨髓浸润、B症状均为影响PTCL预后的因素。结论 PTCL肿瘤组织中VEGF表达和MVD水平较高,这与肿瘤的侵袭性以及疗效和预后相关。     

VEGF在前列腺癌中的表达及与肿瘤MVD的关系

目的:研究前列腺癌组织中的血管内皮生长因子（VEGF）表达与前列腺癌肿瘤微血管密度（MVD）的关系。方法:选取81例确诊前列腺癌患者及良性前列腺增生患者52例。采用免疫组化法进行染色,观察两种组织中VEGF、MVD的表达情况,并分析VEGF与前列腺癌患者的临床病理特征关系及与MVD之间的关系。结果:前列腺癌组织中VEGF阳性表达率（69.14%）显著的高于前列腺良性增生组（28.85%）,差异具有统计学意义（P<0.05）。前列腺癌组织中MVD计数为（36.7±8.2）显著高于前列腺良性增生组（19.3±5.8）,差异具有统计学意义（P<0.05）。前列腺癌组织中VEGF表达阳性率与患者的TNM分期、淋巴结转移、分化程度具有显著关联（P<0.05）。VEGF表达阳性的癌组织中MVD（40.3±7.5）显著多于VEGF表达阴性的癌组织（28.7±6.1）,差异具有统计学意义（P<0.05）。结论:前列腺癌组织中VEGF高表达,MVD生成增多,VEGF与患者的临床病理特征具有一定的关系,VEGF阳性表达患者的MVD增生水平越高。     

喉癌组织中环氧化酶-2表达及与VEGF、MVD的关系

目的　研究喉癌组织中环氧化酶2(COX 2)的表达与临床病理参数及VEGF、MVD的关系,并探讨其临床价值。方法　采用免疫组化技术检测40例喉癌组织中COX 2、VEGF的表达情况,用CD34标记新生血管内皮细胞,在显微镜下观察微血管密度。结果　喉癌组织中COX 2、VEGF的阳性表达率分别为67.5% (27/40)和80% (32/40),MVD平均为52.46±16.96条/200倍视野。与声带息肉及癌周正常组织相比,肿瘤组织中COX 2、VEGF表达及MVD明显增加,COX 2表达与肿瘤临床分期有关。COX 2表达与VEGF有显著相关性,且COX 2、VEGF表达阳性者MVD明显增加。结论　喉癌组织中存在COX 2的高表达。COX 2与肿瘤新生血管形成有关,它可通过增加VEGF的表达促进肿瘤血管形成,从而促进喉癌的生长和转移。     

VEGF及其受体在多发性骨髓瘤中的表达及意义

背景与目的:骨髓新生血管形成在多发性骨髓瘤(multiple myeloma,MM)的发生、发展和预后中起着重要的作用,血管内皮生长因子(vascular endothelialgrowth factor,VEGF)在此过程中扮演了关键角色.本研究旨在研究VEGF及其受体在MM中的表达,分析其与MM发生、发展的关系.方法:采用RT-PCR的方法检测35例MM患者、16例非肿瘤患者(下称对照组)以及KM3细胞株中VEGF及其受体Flt-1和KDR的表达,并分析阳性率以及表达相对含量在MM患者和非肿瘤患者、MM不同分期间的差异.结果:VEGF和Flt-1基因在MM组的阳性率(62.9%和80.0%)显著高于对照组(18.8%和31.3%)(P＜0.01),VEGF在两组中的表达水平为0.41±0.19 vs.0.06±0.01(P＜0.05),Flt-1为0.60±0.33 vs.0.08±0.03(P＜0.01);VEGF基因在初治组和复发/难治MM组的阳性率为66.7%vs.60.9%(P＞0.05),Flt-1基因为83.3%vs.78.3%(P＞0.05);VEGF在Ⅱ期和Ⅲ期MM的阳性率为50%vs.73.7%(P＞0.05),Flt-1为81.3%vs.78.9%(P＞0.05);复发/难治MM组的VEGF和Flt-1基因的表达水平明显高于初治组(0.49±0.20 vs.0.28±0.04,P＜0.05;0.70±0.38 vs.0.41±0.06,P＜0.05),Ⅲ期MM患者VEGF和Flt-1基因的表达水平明显高于Ⅱ期患者(0.48±0.19 vs.0.28±0.09,P＜0.05;0.75±0.35 vs.0.41±0.21,P＜0.05).KDR仅在3例MM患者中检出,对照组未检出.结论:VEGF和Flt-1在MM中高表达,并与疾病进展相关.     

Overexpression of vascular endothelial growth factor (VEGF) and its cellular receptor KDR (VEGFR-2) in the bone marrow of patients with acute myeloid leukemia.

Vascular endothelial growth factor (VEGF) and its cellular receptor VEGFR-2 have been implicated as the main endothelial pathway required for tumor neovascularization. However, the importance of the VEGF/VEGFR-2 system for angiogenesis in hematologic malignancies such as AML remains to be elucidated. In 32 patients with newly diagnosed untreated AML, we observed by immunohistochemical analysis of bone marrow biopsies significantly higher levels of VEGF and VEGFR-2 expression than in 10 control patients (P <0.001). In contrast, VEGFR-1 staining levels in AML patients were in the same range as in the controls. Expression of VEGF and VEGFR-2 was significantly higher in patients with a high degree of microvessel density compared to those with a low degree (VEGF: P =0.024; VEGFR-2: P =0.040) and correlated well with bone marrow microvessel density (r(s)=0.566 and 0.609, respectively; P <0.001). Furthermore, in patients who achieved a complete remission following induction chemotherapy VEGFR-2 staining levels decreased into the normal range. In conclusion, our results provide evidence for increased expression of VEGF/VEGFR-2 of leukemic blasts and correlation with angiogenesis in the bone marrow of AML patients. Thus, VEGF/VEGFR-2 might constitute promising targets for antiangiogenic and antileukemic treatment strategies in AML.     

Increased bone marrow microvascular density in haematological malignancies is associated with differential regulation of angiogenic factors

Antiangiogenic drugs are currently tested in haematological malignancies. As these drugs target different angiogenic regulators, and as cancers are inherently heterogeneous, a detailed characterization of angiogenesis in individual cancers is needed. Hence, we measured bone marrow microvessel density (MVD), plasma concentrations of eight angiogenesis-related parameters and the expression in blood mononuclear cells of 40 angiogenesis-related mRNAs in 93 patients with haematological neoplasias (acute myeloid leukaemia; chronic lymphatic leukaemia; multiple myeloma (MM); or non-Hodgkin's lymphoma (NHL)) before start and after completion of cancer therapy. Compared with healthy individuals, the patients had significantly increased bone marrow MVD, especially patients with advanced stage disease. A novel finding was that patients with NHL also had increased bone marrow MVD. The plasma levels of vascular endothelial growth factor (VEGF), interleukin (IL)-6 and IL-8 were significantly increased. VEGF levels were highest in those who did not achieve complete remission after cancer therapy. The mRNA expression of IL-8 was upregulated 15-fold. Our data show that patients with haematological malignancies have increased bone marrow MVD; hence, supporting the notion that bone marrow angiogenesis plays a role in the pathogenesis and progression of these cancers. VEGF, IL-6 and IL-8 seem to contribute to the malignant phenotype.     

术前化疗对乳腺癌组织VEGF表达和血管生成的影响

目的研究术前化疗对乳腺癌血管内皮生长因子(VEGF)表达、肿瘤组织微血管密度的影响.方法36例术前化疗的乳腺癌和32例未做术前化疗的乳腺癌标本,进行VEGF、CD34免疫组织化学标记,并进行微血管计数和统计学处理.结果术前化疗组VEGF阳性率为47.22%(17/36),对照组53.13%(17/32),微血管计数化疗组和对照组分别为23.67±13.45,26.12±11.32,两者差异均无显著性.化疗组VEGF阳性和阴性的标本,MVC均值差异无显著性;对照组VEGF阳性和阴性的标本,MVC均值差异有显著性.结论常规术前化疗并未显示出明显的血管生成抑制作用,也未显示出明显的血管内皮生长因子表达的改变.     

可溶性细胞间粘附分子-1及血管内皮生长因子在糖尿病合并多发性骨髓瘤患者血清中的表达水平

目的 探讨可溶性细胞间粘附分子-1(sICAM-1)及血管内皮生长因子(VEGF)在糖尿病合并多发性骨髓瘤(MM)患者血清中的表达水平及其关系.方法 随机选择60例MM患者(实验组)以及40例健康体检者(对照组)为研究对象,观察和对比不同临床分期和不同肿瘤量MM患者血清sICAM-1及VEGF的表达水平.结果 健康对照组的sICAM-1、VEGF的表达水平分别为(287±81) μg/L、(113±78) mg/L,实验组患者血清中sICAM-1及VEGF的表达水平则分别为(375±100) μg/L、(614±205) mg/L,与对照组比较,差异有统计学意义(P＜0.05).sICAM-1及VEGF表达水平随着MM肿瘤量的增加和临床分期增高而上升(P＜0.05).MM患者血清中sICAM-1浓度与VEGF浓度呈正相关.结论 sICAM-1及VEGF在MM患者血清中均呈现出高表达情况,且在不同时期MM患者中均有不同程度的增高.因此VEGF及sICAM-1的浓度或许可作为监测MM患者病情的指标之一.     

Serum concentrations and clinical significance of soluble CD40 ligand in patients with multiple myeloma

Multiple myeloma (MM) is a disease of plasma cells that express the CD40 receptor. Binding of the CD40 by its natural ligand, CD40 ligand (CD40L), produces growth arrest and/or apoptosis in MM. To evaluate serum levels of soluble CD40L (sCD40L) in MM patients and to correlate them with markers of disease activity and angiogenesis, such as vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), interleukin-6 (IL-6), proliferation marker Ki-67 proliferation index (Ki-67 PI) and bone marrow plasma cell infiltration, fifty-eight MM patients were studied in diagnosis and 43 of them after completion of treatment. Serum levels of sCD40L, VEGF, HGF and IL-6 were measured by ELISA, whereas Ki-67 PI and bone marrow plasma cell infiltration were measured by immunohistochemistry. Pre-treatment levels of sCD40L in MM patients were higher compared to controls and to their levels after effective treatment. Treatment regimen did not affect the degree of reduction of sCD40L levels, whereas patient in partial remission had increased levels compared to those with better response. Significant differences were found among disease stages. There were also positive correlations between CD40L with HGF, VEGF, IL-6 and Ki-67 PI. Elevated serum sCD40L is found in patients with advanced MM stage and can be reduced after effective treatment. Increased levels of this mediator are correlated with angiogenic cytokines, providing evidences that CD40L/CD40 interactions play a significant role in the mechanisms of angiogenesis in MM patients.     

Predicting treatment responses and disease progression in myeloma using serum vascular endothelial growth factor and hepatocyte growth factor levels

Angiogenesis is a crucial process in the progression of multiple myeloma (MM). Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) are multifunctional cytokines that potently stimulate angiogenesis during tumor neovascularization. VEGF is secreted by MM cells. It induces proliferation of MM cells and stimulates IL-6 expression by microvascular endothelial cells and bone marrow stroma cells, suggesting both autocrine and paracrine functions for VEGF in MM. HGF and the HGF receptor, c-Met, are expressed simultaneously in MM cell lines and in freshly isolated MM cells, suggesting a possible role for HGF in MM cell proliferation. This review focuses on the clinical significance of serum levels of VEGF and HGF in MM.