VEGF-C和VEGF-D在肝内胆管癌组织中的表达与淋巴结转移的关系及临床意义

目的：探讨胆管癌患者血管内皮生长因子C和D（vascular endothelial growth factor-Cand．D,VEGF．CandVEGF．D）在胆管癌组织中的表达及其与肿瘤淋巴结转移的关系。方法：应用免疫组化SABC法及Real-timePCR法检测57例胆管癌组织和正常胆管组织中VEGF-C、vEGF-D蛋白及其mRNA的表达。结果：胆管癌组织VEGF—C和VEGF．D表达明显高于正常胆管组织（P〈0．叭）,其中淋巴结转移组VEGF-C、VEGF—D的表达与淋巴结未转移组间统计学差异显著（P〈0．05）。VEGF-C和VEGF-D在胆管癌组织中的表达与淋巴结转移有关（P〈0．01）。结论：胆管癌细胞非摄入性高表达的VEGF．C和VEGF．D与淋巴结转移密切相关,可作为评估胆管癌患者预后的重要参考指标。     

VEGF-C和VEGF-D在胃癌组织中的表达与微血管生成的关系及临床意义

目的:观察胃癌患者血管内皮生长因子C和D(vascular endothelial growth factor-C and-D,VEGF-C,VEGF-D)在胃癌组织中的表达以及其与肿瘤微血管生成的关系.方法:采用免疫组化S-P法检测30例胃癌组织和30例正常胃组织中VEGF-C和VEGF-D蛋白的表达,检测微血管密度(microvessel density,MVD).结果:胃癌组织VEGF-C和VEGF-D表达明显高于正常胃组织(p<0.01),其中淋巴结转移组VEGF-C、VEGF-D的表达与淋巴结未转移组间差异显著(p<0.05).胃癌组织MVD值明显高于正常胃组织(p<0.01),淋巴结转移组MVD值明显高于未转移组(p<0.01).MVD与VEGF-C的蛋白在胃癌组织中的表达高度相关(r=0.735,p<0.05),与VEGF-D的蛋白在胃癌组织中的表达成正相关(r=0.623,p<0.05).结论:VEGF-C和VEGF-D的高表达与肿瘤微血管生成及淋巴道转移密切相关,可作为评估胃癌患者预后的重要参考指标.     

TrkB在胆管癌中的表达及其临床意义

目的:研究胆管癌和正常胆管组织中TrkB蛋白的表达及其与临床病理特征的关系。方法:免疫组化检测42例胆管癌和10例正常胆管石蜡切片标本中TrkB蛋白的表达并分析其与临床病理因素的关系。结果:正常胆管组织中无TrkB蛋白的表达,胆管癌组织TrkB蛋白的表达率为69.0%(29/42)。TrkB表达与分化程度、有无淋巴结转移和远处转移有关,而与性别、年龄、肿瘤大小无关。TrkB蛋白在低分化组、有淋巴结转移组和有远处转移组中的表达率均高于高中分化组,无淋巴结转移组和无远处转移组(P<0.05)。结论:TrkB在胆管癌组织中的表达可作为预测胆管癌发生淋巴结转移和远处转移的指标。     

VEGF-C 和VEGFR-3 在胃癌组织中的表达及临床意义

目的:探讨胃癌患者血管内皮生长因子C(vascular endothelial growth factor-C,VEGF-C及血管内皮生长因子受体-3(vascular endothelial growth factor receptor-3,VEGFR-3)在胃癌组织中的表达,从而确定胃癌预后的分子标志物。方法:搜集整理临床资料,采用Real-time PCR及ELISA法检测43例胃癌组织VEGF-C和VEGFR-3的表达。结果:43例胃癌组织中均有不同程度的VEGF-C和VEGFR-3的表达,Real-time PCR结果显示胃癌组织淋巴结转移组和非转移组VEGF-C和VEGFR-3的表达分别为0.07±0.01和0.12±0.01,0.03±0.01和0.06±0.02,与正常对照组相比,差异有显著性(p<0.05)。ELISA检测显示,与正常胃组织中VEGF-C和VEGFR-3的蛋白表达相比,胃癌无淋巴结转移组及胃癌并发淋巴结转移组中VEGF-C和VEGFR-3均明显增加。结论:VEGF-C和VEGFR-3的表达与胃癌淋巴结转移密切相关,提示胃癌标本VEGF-C和VEGFR-3的检测可作为胃癌预后的分子标志物。     

胆管癌组织中SNCG的表达及其临床意义(英文)

目的:观察γ-synuclein(SNCG)在胆管癌和正常胆管组织中的表达,并探讨其在胆管癌发生、发展中的临床意义。方法:采用免疫组化方法检测SNCG蛋白在72例胆管癌组织及41例胆管正常组织中的表达水平,并分析其与胆管癌临床病理特征的关系。结果:SNCG蛋白在胆管癌组织中的阳性表达率为73.61%(53/72),高于其在胆管正常组织中的阳性表达率(4.88%,2/41),其差异有统计学意义(P0.01)。SNCG蛋白的表达与肿瘤的淋巴结转移相关(P0.01),但与患者的年龄、性别及肿瘤分化程度无关(P0.05)。结论:SNCG蛋白的表达与胆管癌的发生、发展正相关,并对胆管癌的浸润转移发挥重要的促进作用。对SNCG蛋白的研究将可能为胆管癌的早期诊断提供新的肿瘤标志物,并为胆管癌的预后判断和诊治提供重要理论依据。     

诱导受体3（DcR3）在胆管癌中的表达及意义

目的:观察胆管癌组织及血清中诱导受体3(decoy receptor 3,DcR3)蛋白的表达及其临床价值。方法:采用免疫组化S-P法检测45例胆管癌、15例癌旁胆管正常组织中DcR3蛋白的表达,ELISA法检测31例胆管癌及18例胆道良性疾病患者和28例正常人外周血清中DcR3的水平。结果:45例胆管癌组织中DcR3阳性表达29例,阳性率为64.4%,胆管正常组织中无阳性表达。DcR3的表达与肿瘤临床分期、肿瘤浸润和转移有关(P<0.05)。胆管癌患者及胆管良性疾病患者血清DcR3水平分别为152.2535.94 pg/ml,98.35 14.27 pg/ml,均高于正常人。胆管癌患者与胆道良性疾病患者血清DcR3水平相比差异有显著性(P<0.01)。结论:DcR3在胆管癌组织中表达增高。DcR3的表达与胆管癌的发生、发展以及转移有关,可成为治疗胆管癌的一个新靶点。血清DcR3的检测对胆管癌的诊断有一定的临床价值。     

VEGF-D在甲状腺癌组织中的表达和意义

目的:探讨甲状腺癌中血管内皮生长因子-D(Vascular Endothelial Growth Factor,VEGF-D)在甲状腺癌中的表达及意义.方法:免疫组织化学染色方法分别对16例癌周正常组织和56例甲状腺癌组织切片染色,观察癌周正常组织和甲状腺癌组织VEGF-D的表达情况.结果:(1)在甲状腺癌中VEGF-D阳性表达主要见于细胞的胞膜上和胞质中.VEGF-D阳性表达在癌周正常组织和甲状腺癌分别为18.7%和71.4%,两者在统计学上有显著差异.(2)在甲状腺癌中VEGF-D在淋巴结非转移组表达阳性率为60.0%,而转移组表达为83.8%,明显高于非转移组,两者问在统计学上有显著差异.(3)在甲状腺乳头状癌中VEGF-D阳性表达在非转移组50%,淋巴结转移组85.1%,明显高于非转移组,说明VEGF-D在淋巴结转移组比非淋巴结转移组表达显著,VEGF-D的阳性表达与甲状腺癌的淋巴结转移显著相关.结论:(1)在癌周甲状腺组织VEGF-D阳性反应较弱,在甲状腺癌组织VEGF-D阳性反应较强,两者比较有显著性差异.(2)在甲状腺癌中,尤其是甲状腺乳头状癌中,其癌组织中VEGF-D的表达强度增加,与无淋巴结转移的甲状腺癌组织相比,有淋巴结转移的甲状腺癌组织中VEGF-D的表达较强,可作为肿瘤转移的指标之一.     

VEGF-D和CD105在胰腺癌中的表达及意义

目的:探讨VEGF-D,CD105在胰腺癌发生、发展及转移过程中的意义.方法:采用免疫组织化学S-P法,检测VEGF-D,CD105在53例胰腺癌组织(胰腺癌组)及10例正常胰腺组织(对照组)中的表达,计数MVD,并与临床病理学资料进行分析比较.结果:胰腺癌组织中VEGF-D的阳性表达明显高于正常胰腺组织(P＜0.05).VEGF-D的表达与胰腺癌的临床分期及淋巴结有无转移有关(P＜0.05).胰腺癌中,CD105标记的微血管密度(MVD)明显高于非肿瘤性胰腺组织.CD105的表达水平与胰腺癌临床分期及病理分级有关.胰腺癌组织中CD105抗体标记的微血管密度(MVD)结果显示:在有淋巴结转移的癌组织中,其MVD值高于无淋巴结转移组有显著统计学意义(P＜0.05).VEGF-D阳性表达组的MVD显著高于VEGF-D表达阴性组的MVD(P＜0.05):CD105在胰腺癌中的表达与VEGF-D的表达呈正相关(P＜0.05).结论:VEGF-D的表达与胰腺癌的血管生成有关.MVD值有可能作为判断胰腺癌生物学行为的潜在指标.     

VEGF-C和CCR7的表达与卵巢癌淋巴结转移之间的关系

目的观察血管内皮生长因子(VEGF)-C和趋化因子受体CCR7在卵巢癌组织内的表达情况,分析VEGF-C和CCR7的表达与癌淋巴结转移之间的关系。方法取卵巢癌病例72例,其中,淋巴结转移组46例,无淋巴结转移组26例。应用免疫组化技术观察VEGF-C和CCR7在卵巢癌组织内的表达。结果 VEGF-C和CCR7主要表达于卵巢癌细胞胞浆或/和胞膜内,VEGF-C和CCR7在淋巴结转移组的阳性表达率分别是71.7%和78.2%,在无淋巴结转移组的表达率分别是30.8%和26.9%,二者在淋巴结转移组的表达率均明显高于无淋巴结转移组(P<0.01)。VEGF-C和CCR7蛋白同时阳性表达在淋巴结转移组和非淋巴结转移组中的表达率分别为65.2%和15.4%,VEGF-C和CCR7的表达具有显著的相关性(P<0.01),联合检测VEGF-C和CCR7诊断卵巢癌淋巴结转移具有较高的准确度,ROC曲线下面积达0.791。结论 VEGF-C和CCR7表达在卵巢癌淋巴结转移过程中发挥重要作用,VEGF-C和CCR7在促进卵巢癌淋巴结转移中可能具有一定的协同作用,二者联合检测有助于预示卵巢癌淋巴结转移的判断。     

VEGF-C 在口腔鳞状细胞癌中的表达及其意义

目的:检测血管内皮生长因子-C(VEGF-C)在口腔鳞状细胞癌(鳞癌)和癌旁组织中的表达情况,探讨VEGF-C在口腔鳞癌的增殖、浸润和淋巴转移中的作用。方法:采用免疫组化方法检测60例口腔鳞癌病人癌组织和癌旁组织中VEGF-C的表达,应用图像分析系统进行分析,用Spearman相关分析研究其与病变部位、肿瘤大小、病理分级、临床分期及颈淋巴结转移之间的关系。结果:口腔鳞癌组织VEGF-C的表达明显高于癌旁组织(u=7.747,P<0.01),其表达强度与临床分期及淋巴结转移密切相关(r=0.564、0.706,P<0.05),与病变部位、大小、病理分级无关。结论:口腔鳞癌细胞分泌VEGF-C诱导癌周淋巴管增生扩张是发生区域淋巴结转移的重要因素之一,VEGF-C有望作为早期临床判断和预测颈淋巴结转移的指标之一。     

上皮性卵巢癌中Maspin蛋白与血管内皮生长因子-C的表达及其相关性

目的：探讨上皮性卵巢癌组织中maspin蛋白与血管内皮生长因子-C（VEGF-C）表达的临床意义及其相关性。方法：采用免疫组化技术检测75例上皮性卵巢癌中maspin蛋白与VEGF-C的表达情况,以卵巢良性肿瘤及正常卵巢作为对照。结果：maspin和VEGF-C在上皮性卵巢癌中的阳性表达率分别为61.3%、82.7%,均明显高于卵巢良性肿瘤（13.3%、20%）和正常卵巢组织（0%、0%）,maspin蛋白在上皮性卵巢癌中的表达与FIGO分期高、组织学分级高、腹水形成和淋巴结转移有关;VEGF-C与FIGO分期高、淋巴结转移和腹水形成有关;上皮性卵巢癌中maspin蛋白与VEGF-C的表达成正相关。结论：maspin和VEGF-C在上皮性卵巢癌中表达上调,在卵巢上皮性癌的浸润、转移中起重要作用。     

VEGF-C和VEGFR-3在胃癌组织中的表达及临床意义

目的：探讨胃癌患者血管内皮生长因子C（vascular endothelial growth factor-C,VEGF-C及血管内皮生长因子受体-3（vascular endothelial growth factor receptor-3,VEGFR-3）在胃癌组织中的表达,从而确定胃癌预后的分子标志物。方法：搜集整理临床资料,采用Real-time PCR及ELISA法检测43例胃癌组织VEGF-C和VEGFR-3的表达。结果：43例胃癌组织中均有不同程度的VEGF-C和VEGFR-3的表达,Real-time PCR结果显示胃癌组织淋巴结转移组和非转移组VEGF-C和VEGFR-3的表达分别为0.07±0.01和0.12±0.01,0.03±0.01和0.06±0.02,与正常对照组相比,差异有显著性（p〈0.05）。ELISA检测显示,与正常胃组织中VEGF-C和VEGFR-3的蛋白表达相比,胃癌无淋巴结转移组及胃癌并发淋巴结转移组中VEGF-C和VEGFR-3均明显增加。结论：VEGF-C和VEGFR-3的表达与胃癌淋巴结转移密切相关,提示胃癌标本VEGF-C和VEGFR-3的检测可作为胃癌预后的分子标志物。     

Restricted expression pattern of vegf-d in the adult and fetal mouse: high expression in the embryonic lung

Endothelial growth factors have become the target of intense research since the initial discovery of vascular endothelial growth factor (VEGF/VPF). At present, VEGF is established as a major inducer of angiogenesis in normal and pathological conditions. Recently several new members in the VEGF family have been described; VEGF-B/VRF, VEGF-C and VEGF-D. VEGF-D is most closely related to VEGF-C by virtue of the presence of N- and C-terminal extensions that are not found in other VEGF family members. We have here examined the expression pattern of vegf-d mRNA with in situ hybridization in developing and adult mice. This shows a restricted expression pattern, with high levels mainly in lung tissue. The expression in embryonic lung is upregulated prior to birth. Expression of vegf-d in other tissues, as well as in lung tissue of the E14 embryo, was either low or absent. This suggests that VEGF-D may be of special relevance for the vascularization of lung tissue during the last trimester of fetal development.     

Differential mRNA and tissue expression of lymphangiogenic growth factors (VEGF-C and -D) and their receptor (VEGFR-3) during tail regeneration in a gecko

Lymphangiogenesis, the growth of new lymph vessels, has important roles in both normal and pathological lymphatic function. Despite recent advances, the precise molecular mechanisms behind the lymphangiogenic process remain unclear. The Australian marbled gecko, Christinus marmoratus, voluntarily drops its tail (autotomy) as a predator avoidance strategy. Following autotomy a new tail is regenerated including lymphatic drainage pathways. We examined the molecular control of lymphangiogenesis within the unique model of the regenerating gecko tail. Partial sequences were obtained of the gecko lymphangiogenic growth factors, vascular endothelial growth factor C (VEGF-C) and VEGF-D along with their receptor VEGFR-3. These were highly homologous to other vertebrates. Quantitative real-time polymerase chain reaction (PCR) demonstrated up-regulation of VEGF-C, VEGF-D and VEGFR-3 mRNA expression during the early and middle stages of tail regeneration (between 4 and 9 weeks following autotomy), in late regeneration (12 weeks) and during mid-regeneration (7 and 9 weeks), respectively. VEGF-C and VEGF-D immunostaining was observed lining some lymphatic-like and blood vessels in early–mid tail regeneration, indicating possible associations of the proteins with VEGFRs on endothelia. Keratinocytes and fibroblasts also showed positive staining of VEGF-C and VEGF-D in early–mid tail regeneration. Additionally, VEGF-C was localised in adipose tissue in all tail states examined. This work suggests that specific timings exist for the expression of the lymphangiogenic growth factors, VEGF-C and VEGF-D, and their receptor, VEGF-R3, throughout the regeneration of a functional lymphatic network. Along with localisation data, this suggests potential functions for the growth factors in lymphangiogenesis and angiogenesis throughout tail regeneration.     

Expression of vascular endothelial growth factors VEGF- C and D, VEGFR-3, and comparison of lymphatic vessels density labeled with D2-40 antibodies as a prognostic factors in vulvar intraepithelial neoplasia (VIN) and invasive vulvar cancer

OBJECTIVE: The aim of this study was to compare the immunohistochemical expression of vascular endothelial growth factors VEGF-C and D, as well as the expression of VEGFR-3 in VIN and vulvar invasive cancer and to compare the density of lymphatic marker D2-40 antibody in both groups, and to compare them with different clinicopathologic features. Materials & Methods: The study was performed using tissue material and clinical data from 100 women diagnosed with VIN and 100 women diagnosed with invasive vulvar cancer. Results: No significant differences were found in the expression of VEGF-C and -D or VEGFR-3 between those patients with VIN and those with invasive vulvar cancers. Weak expression of VEGF-C was confirmed only in two cases of the analyzed series; in all cases, expression of VEGF-D and VEGFR-3 was observed. The strongest expression of VEGF-D and VEGFR-3 was observed in the group of invasive cancers. The highest density of lymphatic vessels per 2 mm was observed in VIN. In the cancer group, small lymphatic vessels with a narrow oval lumen were observed. Moreover, in two cases of vulvar cancer, the presence of intratumoral lymphatic vessels was observed. Conclusions: These results suggest that lymphangiogenesis begins at the preinvasive stage of vulvar carcinogenesis and suggests the important role of VEGF-C, VEGF-D, VEGFR-3 and LV (D2-40) as prognostic factors in the process of carcinogenesis in the vulvar area.     

Localisation of lymphatic vessels and vascular endothelial growth factors-C and -D in human and mouse skeletal muscle with immunohistochemistry

The present study was aimed to localise lymphatic vessels and their growth factors in human and mouse skeletal muscle with immunohistochemistry and specific antibodies (VEGFR-3, LYVE-1, VEGF-C and VEGF-D). The largest lymphatic vessels were found in perimysial connective tissue next to the arteries and veins, as has been shown earlier with electron microscopy. As a new finding, we also found small LYVE-1 positive vessels in the capillary bed between muscle fibres. These vessels were located next to CD31 positive blood capillaries and were of the same size, but fewer in number. In addition, we described the localisation of the two main lymphangiogenic growth factor proteins, vascular endothelial growth factor-C and -D. Both proteins were expressed in skeletal muscle at mRNA and protein levels. VEGF-D was located under the sarcolemma in some of the muscle fibres, in the endothelia of larger blood vessels and in fibroblasts. VEGF-C protein was localised to the nerves and muscle spindles, to fibroblasts and surrounding connective tissue, but was not found in muscle fibres or endothelial cells. Our results are the first to suggest the presence of lymphatic capillaries throughout the skeletal muscle, and to present the localisation of VEGF-C and -D in the muscles. Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

Characterization of indolinones which preferentially inhibit VEGF-C- and VEGF-D-induced activation of VEGFR-3 rather than VEGFR-2.

VEGF-C and VEGF-D are lymphangiogenic factors that bind to and activate VEGFR-3, a fms-like tyrosine kinase receptor whose expression is limited almost exclusively to lymphatic endothelium in the adult. Processed forms of VEGF-C and VEGF-D can also activate VEGFR-2, a key player in the regulation of angiogenesis. There is increasing evidence to show that these receptor-ligand interactions play a pivotal role in a number of pathological situations. Inhibition of receptor activation by VEGF-C and VEGF-D could therefore be pharmaceutically useful. Furthermore, to understand the different roles of VEGF-C, VEGF-D, VEGFR-2 and VEGFR-3 in pathological situations it will be necessary to dissect the complex interactions of these ligands and their receptors. To facilitate such studies we cloned, sequenced and characterized the expression of rat VEGF-C and VEGF-D. We showed that Cys152-->Ser mutants of processed rat VEGF-C can activate VEGFR-3 but not VEGFR-2, while the corresponding mutation in rat VEGF-D inhibits its ability to activate both VEGFR-2 and VEGFR-3. We also synthesized and characterized indolinones that differentially block VEGF-C- and VEGF-D-induced VEGFR-3 kinase activity compared to that of VEGFR-2. These tools should be useful in analysing the different activities and roles of VEGF-C, VEGF-D and their ligands, and in blocking VEGFR-3-mediated lymphangiogenesis.     

Vascular endothelial growth factor-D is a key molecule that enhances lymphatic metastasis of soft tissue sarcomas

Studies on lymph node metastasis of soft tissue sarcomas are insufficient because of its rarity. In this study, we examined the expressions of vascular endothelial growth factor (VEGF)-C and VEGF-D in soft tissue sarcomas metastasized to lymph nodes. In addition, the effects of the two molecules on the barrier function of a lymphatic endothelial cell monolayer against sarcoma cells were analyzed. We examined 7 patients who had soft tissue sarcomas with lymph node metastases and who had undergone neither chemotherapy nor radiotherapy before lymphadenectomy. Immunohistochemistry revealed that 2 of 7 sarcomas that metastasized to lymph nodes expressed VEGF-C both in primary and metastatic lesions. On the other hand, VEGF-D expression was detected in 4 of 7 primary and 7 of 7 metastatic lesions, respectively. Interestingly, 3 cases that showed no VEGF-D expression at primary sites expressed VEGF-D in metastatic lesions. Recombinant VEGF-C at 10(-8) and VEGF-D at 10(-7)and 10(-8)g/ml significantly increased the random motility of lymphatic endothelial cells compared with controls. VEGF-D significantly increased the migration of sarcoma cells through lymphatic endothelial monolayers. The fact that VEGF-D induced the migration of fibrosarcomas through the lymphatic endothelial monolayer is the probable reason for the strong relationship between VEGF-D expression and lymph node metastasis in soft tissue sarcomas. The important propensities of this molecule for the increase of lymph node metastases are not only lymphangiogenesis but also down-regulation of the barrier function of lymphatic endothelial monolayers, which facilitates sarcoma cells entering the lymphatic circulation.