血管内皮生长因子受体3和Podoplanin在人结肠癌组织中的表达

目的:观察血管内皮生长因子受体3(vascular endothelial growth factor receptor-3,VEGFR-3)和Podoplanin在人结肠癌组织中的表达,以探讨二者标记淋巴管的特异性。方法:选择55例人结肠癌组织标本,应用免疫组织化学法观察VEGFR-3和Podoplanin在人结肠癌组织中的表达。结果:Podoplanin主要表达于淋巴管内皮细胞上,微血管极少着色;VEGFR-3主要表达于淋巴管内皮细胞上,另外在小血管内皮也有较丰富的表达。结论:Podoplanin在淋巴管内皮细胞的表达具有较高特异性,可以用来标记淋巴管。     

人喉癌组织中VEGF-C、VEGF-D及其受体3的表达及意义

目的 探讨喉癌组织中VEGF—C、D和受体VEGFR-3的表达及其在喉癌进展中的作用。方法 取人喉癌标本12例,正常及良性病变喉组织10例,免疫组化法观察VEGF—C、VEGF—D、VEGFR-3以及LYVE-1的表达。结果 VEGF—C和VEGF—D主要表达于喉癌细胞胞浆内,喉癌组织中VEGF—C和VEGF—D表达的阳性率明显高于正常及良性病变喉组织（P〈0．05）;VEGFR-3主要表达于基底层的癌细胞,在喉癌组织中VEGFR-3表达的阳性率明显高于正常和良性病变组织中（P〈0．01）,并且VEGFR-3的表达与VEGF—C、VEGF—D的表达显著正相关（P〈0．01）。LYVE—1仅见表达于淋巴管内皮细胞。结论 喉癌组织中VEGF—C、VEGP-D的表达明显增高,推测可能通过与VEGFR-3的结合促进喉癌组织中淋巴管的生成;LYVE-1是淋巴管内皮细胞较特异的标记物。     

血管内皮生长因子C及其受体3在人恶性黑色素瘤组织内的表达及意义

目的 观察人恶性黑色素瘤组织内血管内皮生长因子C(VEGF-C)及其受体3(VEGFR-3)的表达,探讨VEGF-C和VEGFR-3在恶性黑色素瘤淋巴管生成及淋巴道转移中的作用.方法 取人恶性黑色素瘤组织48例(石蜡标本30例,术后新鲜组织18例),应用免疫组织化学和RT-PCR技术,观察VEGF-C和VEGFR-3蛋白及mRNA在恶性黑色素瘤组织内的表达情况.以淋巴管内皮透明质酸受体(LYVE-1)标记淋巴管,计数恶性黑色素瘤组织淋巴管数密度.结果 VEGF-C和VEGFR-3蛋白主要表达于恶性黑色素瘤细胞胞浆内,在肿瘤周围的血管和淋巴管内皮上也可见VEGFR-3蛋白表达,VEGF-C和VEGFR-3蛋白在淋巴结转移组恶性黑色素瘤组织内的表达水平明显高于无淋巴结转移组(P<0.05).在18例新鲜恶性黑色素瘤中,淋巴结转移组VEGF-C和VEGFR-3mRNA的表达明显高于无淋巴结转移组(P<0.01).LYVE-1表达于肿瘤间质内的淋巴管内皮细胞,淋巴结转移组恶性黑色素瘤组织中的淋巴管数密度(LMVD)为9.845±2.454,无淋巴结转移组恶性黑色素瘤组织中的淋巴管数密度为6.534±2.193,淋巴结转移组恶性黑色素瘤组织内的淋巴管数密度明显高于无淋巴结转移组(P<0.01).结论恶性黑色素瘤组织内VEGF-C表达明显增高,并通过上调其受体VEGFR-3的表达促进恶性黑色素瘤组织内淋巴管的生成,从而促进恶性黑色素瘤的淋巴道转移.     

胃癌组织淋巴管LYVE-1、钙粘蛋白的表达及其与癌转移的关系

目的：探讨胃癌组织钙粘蛋白的表达与癌淋巴道转移的相关性。方法：取胃癌标本14例，正常胃组织标本2例。通过E-cadherin、β-catenin和LYVE-1免疫组化方法，观察肿瘤淋巴管的表达。结果：LYVE-1在正常和癌组织淋巴管阳性表达，正常淋巴管E．cadherin、β-catenin表达阴性，β-catenin在胃癌淋巴管表达弱阳性，β-catenin在淋巴管的表达与肿瘤的分化程度及有无转移有负相关性。结论：LYVE-1在淋巴管特异性表达，E-cadherin／β-catenin复合物在淋巴道转移过程中，对肿瘤细胞与淋巴管内皮细胞的粘附起一定的作用，但不是主要作用。     

血管内皮生长因子C及其受体在MNNG诱发的大鼠大肠癌细胞及淋巴管的表达

目的 观察大鼠大肠癌组织内血管内皮生长因子-C(VEGF-C)及其受体3(VEGFR-3)的表达情况,探讨VEGF-C及其受体VEGFR-3在肿瘤淋巴转移中的作用.方法 采用甲基硝基亚硝基胍(MNNG)诱发的大鼠大肠癌模型,应用免疫组织化学(SABC法)技术检测29例大鼠原发性大肠癌组织中VEGF-C及VEGFR-3蛋白,观察VEGF-C及VEGFR-3在大肠癌组织内的表达.结果 正常大肠组织内未见VEGF-C阳性表达,但可见淋巴管内皮细胞VEGFR-3阳性表达.在大肠癌组织内,VEGF-C蛋白表达于癌细胞,早期和中晚期癌的阳性表达率分别是75%和100%,(P＜0.05).VEGFR-3主要表达于淋巴管内皮细胞,早期和中晚期癌组织内淋巴管的阳性表达率分别是58.33%和94.12%(P＜0.05).结论 大鼠大肠癌VEGF-C的表达与肿瘤进展有关,推测VEGF-C通过受体VEGFR-3诱导淋巴管生成:VEGFR-3在淋巴管的阳性表达均随肿瘤进展增高,可能与大肠癌淋巴转移有关.     

小鼠移植瘤淋巴管分布及细胞间粘附分子-1的表达

目的：观察小鼠移植瘤内淋巴管的分布及细胞间粘附分子（ICAM-1）在癌细胞和淋巴管内皮细胞的表达。方法：于小鼠腹股沟区皮下接种肝癌细胞株（H22），分期取材。用5＇-Nase-Alpase双重染色法和VEGFR-3免疫组化法观察淋巴管的分布；检测ICAM-1在癌细胞和淋巴管内皮细胞的表达。结果：在肿块的周边部可见少量褐色的毛细淋巴管，VEGFR-3阳性表达。ICAM-1在肿瘤细胞和淋巴管内皮细胞都有表达，随肿瘤的发展而增强。结论：在移植瘤中存在毛细淋巴管，可能为新生的；ICAM-1在肿瘤细胞及淋巴管内皮细胞的表达，可能与癌淋巴管转移有关。     

LYVE-1蛋白在小鼠碱烧伤角膜的表达及意义

目的观察淋巴管内皮透明质酸受体(LYVE-1)在小鼠碱烧伤角膜内的表达情况,探讨角膜新生淋巴管形成的时间过程及角膜疾病后新生淋巴管形成的作用。方法应用NaOH溶液制作小鼠角膜碱烧伤模型,分别于角膜碱烧伤后第1d、3d、5d、7d和12d取材。采用免疫组化法,观察正常角膜和碱烧伤后不同时间段角膜内LYVE-1的表达情况。结果在正常角膜组织中,LYVE-1表达于角膜上皮细胞和内皮细胞内。在角膜碱烧伤后1d、3d和5d,LYVE-1主要表达于角膜上皮内及入侵角膜基质的炎性细胞内;碱烧伤后7d,可见大量LYVE-1呈条索样表达于角膜基质中,并可见少量LYVE-1阳性表达于开放状态的淋巴管;碱烧伤后12d,角膜基质内新生淋巴管数量增多。结论正常角膜组织储备LYVE-1生物因子,在炎性角膜中,LYVE-1可能在角膜新生淋巴管运输透明质酸(HA)的过程中发挥重要作用。     

食管癌血管内皮生长因子和受体的表达及在癌转移中的作用

目的观察人食管癌组织和淋巴管中血管内皮生长因子(VEGF-C)及其受体-3(VEGFR-3)的表达,探讨食管癌的淋巴道转移机制。方法取临床手术切除的食管癌组织,用免疫组化方法检测人食管癌早期和进展期癌细胞或淋巴管对VEGF-C及其VEGFR-3的表达情况。结果在食管癌的癌细胞中可见VEGF-C阳性表达,阳性颗粒主要定位于肿瘤细胞胞浆内。淋巴管内皮细胞仅见VEGFR-3阳性表达,VEGFR-3在血管和癌细胞中也存在少量阳性表达。进展期食管癌VEGF-C和VEGFR-3的表达率和表达强度均强于早期。结论食管癌癌细胞VEGF-C的表达和淋巴管内皮细胞上VEGFR-3的表达均与肿瘤进展呈正相关,推测VEGF-C通过受体VEGFR-3促进食管癌组织淋巴管生成,从而引起癌淋巴道转移。     

淋巴管新生及其在相关疾病发生和治疗中的意义

淋巴管在维持体内微环境衡定、免疫反应和肿瘤淋巴转移等方面起着重要作用,淋巴管新生与胚胎发育、外伤修复、炎症转归和肿瘤转移密切相关.在趋化因子和生长因子的作用下,淋巴管内皮细胞迁移、增殖和构成管腔,形成新的淋巴管.近年来发现淋巴管内皮祖细胞参与淋巴管新生.淋巴管内皮特异表达Prox-1、podoplanin、VEGFR-3和 LYVE-1等,这些因子和受体调控淋巴管新生.VEGF-C/VEGFR-3或VEGF-D/VEGFR-3信号途径在淋巴管新生过程中起着重要作用.VEGF-C、VEGF-D和VEGFR-3可作为基因治疗的靶点,有望治疗淋巴管新生障碍性疾病以及抗移植后免疫排斥和抗肿瘤淋巴管转移.本文主要综述了胚胎发育、先天性淋巴水肿、炎性病变和肿瘤等状态下淋巴管新生的变化及其机制、淋巴管内皮祖细胞参与淋巴管新生的过程、淋巴管内皮细胞特异性转录因子和受体对于淋巴管新生的调控作用.     

乳腺癌淋巴管LYVE-1和PROX-1表达程度与肿瘤淋巴转移的关联性分析

目的观察乳腺浸润性导管癌患者肿瘤组织中乳腺癌淋巴管LYVE-1和PROX-1的表达程度与肿瘤淋巴转移的关联性。方法选择乳腺浸润性导管癌患者90例为实验组,以60例乳腺良性病变患者为对照组,检测实验组患者肿瘤组织LYVE-1和PROX-1的水平,与对照组患者组织的LYVE-1和PROX-1的水平进行比较;将实验组患者按转移、复发情况进行分类,观察LYVE-1和PROX-1表达与肿瘤淋巴转移的关系。结果实验组患者肿瘤组织的LYVE-1和PROX-1阳性淋巴管密度(LVD)明显高于对照组,实验组患者中有淋巴转移患者的LYVE-1和PROX-1阳性LVD明显高于无淋巴转移患者,差异具有统计学意义(P<0.05);LYVE-1和PROX-1阳性LVD之间无相关性;logistic回归分析,LYVE-1和PROX-1是肿瘤转移的相关因素,对LYVE-1、PROX-1与转移的关系绘制ROC曲线,AUC分别为0.716、0.672;按ROC曲线分析所得诊断临界值进行分组,随访观察一年,LYVE-1、PROX-1阳性LVD升高患者,淋巴转移率较高。结论乳腺浸润性导管癌患者肿瘤组织LYVE-1和PROX-1表达与淋巴管生成有关,表达程度高的患者更易发生淋巴转移。     

Immunohistochemical detection of human small lymphatic vessels under normal and pathological conditions using the LYVE-1 antibody

The spread of tumor cells via lymphatic vessels to the lymph nodes is an important indicator of malignancy. However, previous markers used to identify lymphatic endothelium gave ambiguous results in immunohistochemical analyses with paraffin-embedded tissues. In this study, we attempted to prepare a polyclonal antibody against human lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) for detecting lymphatic vessels using immunohistochemistry. The antibody was raised against a region near the transmembrane anchor of LYVE-1 in New Zealand white rabbits. Immunostainings with anti-LYVE-1 and von Willebrand factor antibodies were performed in various normal and pathological tissues. LYVE-1 expression was confined to the endothelial surface of lymphatic vessels but was not found in the endothelium of blood vessels, which were positive for von Willebrand factor. Our LYVE-1 polyclonal antibody was useful for the identification of small lymphatic vessels in normal human tissues. In addition, the immunostaining enabled us to distinguish lymphatic invasion by malignant tumor cells from blood vessel invasion using paraffin-embedded sections. In conclusion, our polyclonal antibody against the transmembrane anchor of the peptide can be used to detect human lymphatic vessels under various conditions.     

肺癌组织及淋巴管E-cadherin、β-catenin和LYVE-1的表达

目的观察肺癌组织E-cadherin和β-catenin的表达,LYVE-1特异性标记淋巴管;探讨钙黏蛋白及其受体在癌细胞淋巴道转移中的作用。方法取肺癌手术材料30例,通过免疫组化法,观察E-cadherin、β-catenin和LYVE-1在癌细胞及淋巴管的表达。结果癌细胞对E-cadherin、β-catenin呈阳性表达,低分化组、有淋巴结转移组E-cadherin表达减弱。淋巴管对LYVE-1阳性表达,E-cadherin阴性表达,β-catenin弱阳性表达。结论LYVE-1在淋巴管特异性表达;E-cadherin表达与肿瘤分化程度和有无淋巴结转移呈负相关;E-cadherin/β-catenin复合物对肿瘤细胞与淋巴管内皮细胞的黏附不起主要作用。     

人胰腺癌淋巴管的分布及形态观察

目的观察人胰腺癌淋巴管的分布及形态结构,探讨胰腺癌淋巴道转移机制。方法取手术后人胰腺癌标本21例,应用免疫组化染色法LYVE-1标记淋巴管进行淋巴管计数,半薄切片光镜观察和超薄切片透射电镜观察胰腺癌组织淋巴管的形态及分布特点。结果胰腺癌组织中LYVE-1染色阳性的脉管具有淋巴管的形态学特征,可见癌周组织的微淋巴管数量较癌旁"正常区"有所增加(P<0.01);半薄切片光镜下可见癌周边区和"正常区"淋巴管存在,癌中心区未见有淋巴管;电镜下癌周边区淋巴管内皮细胞连接开放,部分内皮细胞破裂溶解,管壁不完整。淋巴管内皮细胞的线粒体、高尔基体等细胞器改变。结论胰腺癌组织淋巴管主要位于癌周围浸润区的纤维结缔组织中,且淋巴管数量较癌旁"正常区"增多,淋巴管内皮超微结构改变。胰腺癌淋巴管转移可能通过增多的淋巴管的内皮连接开放和对内皮细胞的破坏溶解作用进入淋巴管管壁。     

VEGFR-3 and Its Ligand VEGF-C Are Associated with Angiogenesis in Breast Cancer

Recently, monoclonal antibodies against the human vascular endothelial growth factor receptor VEGFR-3 were shown to provide a specific antigenic marker for lymphatic endothelium in various normal tissues. In this study we have investigated the expression of VEGFR-3 and its ligand VEGF-C in normal breast tissue and in breast tumors by immunohistochemistry. VEGFR-3 was weakly expressed in capillaries of normal breast tissue and in fibroadenomas. In intraductal breast carcinomas, VEGFR-3 was prominent in the "necklace" vessels adjacent to the basal lamina of the tumor-filled ducts. VEGF receptor 1 and 2 as well as blood vessel endothelial and basal lamina markers were colocalized with VEGFR-3 in many of these vessels. Antibodies against smooth muscle alpha-actin gave a weak staining of the necklace vessels, suggesting that they were incompletely covered by pericytes/smooth muscle cells. A highly elevated number of VEGFR-3 positive vessels was found in invasive breast cancer in comparison with histologically normal breast tissue (P < 0.0001, the Mann-Whitney test). VEGF-C was located in the cytoplasm of intraductal and invasive cancer cells. The results demonstrate that the expression of VEGFR-3 becomes up-regulated in the endothelium of angiogenic blood vessels in breast cancer. The results also suggest that VEGF-C secreted by the intraductal carcinoma cells acts predominantly as an angiogenic growth factor for blood vessels, although this paracrine signaling network between the cancer cells and the endothelium may also be involved in modifying the permeabilities of both blood and lymphatic vessels and metastasis formation.     

VEGF-D in association with VEGFR-3 promotes nodal metastasis in human invasive lobular breast cancer

We assessed the expression of vascular endothelial growth factors (VEGF-C and VEGF-D) in breast cancer cells and the density of lymph vessels and VEGF receptor-3 (VEGFR-3)-positive vessels in and around the tumor in invasive lobular breast cancer. We found significant correlation between peritumoral lymph vessel density and presence of lymph node metastases (P=.001) and the number of metastatic lymph nodes (P<.001). A significant correlation was detected between tumor cell VEGF-D expression and lymph node status (P=.001) and density of lymphatic vessel endothelial receptor (LYVE)-1-positive vessels (P=.035). VEGFR-3+/VEGF-D+ and VEGFR-3+/VEGF-C+ tumors had a significantly higher number of metastatic lymph nodes than tumors with other staining patterns (P<.001). Tumors positive for neither VEGF-D nor VEGFR-3 had a lower density of LYVE-1+ vessels than tumors with other staining patterns (P=.033). Our results indicate that peritumoral lymph vessel density is associated with lymph node metastases in invasive lobular breast cancer and that invasive lobular cancer producing VEGF-D, surrounded by VEGFR-3+ vessels, has a significantly higher peritumoral lymph vessel density and a higher number of metastatic lymph nodes.     

CD40 ligation induces tissue factor expression in human vascular smooth muscle cells

Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are important regulators of blood and lymphatic vessel growth and vascular permeability. Both blood and lymphatic vessels of the upper respiratory tract play important roles in pathological conditions, such as infections and tumors. Here we have studied the expression of VEGF-C and its receptor VEGFR-3 in the upper respiratory system by Northern blot analysis and immunohistochemistry of human tissues, and in situ mRNA hybridization of developing mouse embryos and β-galactosidase staining of mouse embryos having a LacZ marker gene in the VEGFR-3 gene locus. The results demonstrate expression of VEGF-C and VEGFR-3 in the developing and adult nasal respiratory epithelium and in the nasal vascular plexus, respectively. Unlike in most other tissues, in the nasal mucosa VEGFR-3 is expressed in both blood and lymphatic vessels. Expression of VEGF-C was also detected in nasal and nasopharyngeal tumor islands, which were surrounded by VEGFR-3-positive angiogenic blood vessels. These results suggest that VEGF-C and VEGFR-3 have a role in the development of the nasal submucosal vascular plexus and in its normal function and that they are associated with angiogenesis in nasal and nasopharyngeal tumors.     

E-钙黏蛋白、β-连环蛋白和Podoplanin在人胃腺癌中的表达及意义

目的：探讨E-钙黏蛋白（E—cadhefin）及β-连环蛋白（β-catenin）在人胃腺癌中的表达和在淋巴道转移中的作用。方法：取60例胃腺癌组织样本和30例正常组织,应用免疫组织化学方法观察E—Cad和β-Cat在人胃腺癌组织中的表达,应用Podoplanin标记淋巴管,检测胃腺癌组织中的微淋巴管密度（MLVD）。结果：60例胃腺癌组织中,E—Cad和β—Cat的表达与分化程度、淋巴结转移、TNM分期具有相关性,差异有统计学意义（P〈0．05）。E—cad的表达与肿瘤直径、浸润深度具有相关性,差异有统计学意义（P〈0．05）,而β-Cat的异常在这两组中无明显差异（P〉0．05）。Podoplanin只表达淋巴管,癌组织中的LMVD高于正常胃组织（P〈0．01）,癌周边缘区的LMVD高于肿瘤中心组织（P〈0．01）。LMVD的表达与分化程度、淋巴结转移、TNM分期具有相关性,差异有统计学意义（P〈0．05）。结论：E—Cad和β—Cat异常表达在胃腺癌进展中起重要作用,在淋巴转移过程中使肿瘤细胞进入淋巴导管起了一定促进作用。