胃癌淋巴管生成与淋巴结转移的关系

目的:研究胃癌淋巴管生成与淋巴结转移的关系。方法:应用逆转录-聚合酶链反应(RT-PCR)和免疫细胞化学方法检测血管内皮生长因子-C(VEGF-C)和血管内皮生长因子受体-3(VEGFR-3)mRNA及其蛋白在5株胃癌细胞株和3对伴有淋巴结转移的胃癌组织及相应的正常黏膜中的表达。此外,还应用免疫组织化学方法检测86例胃癌标本的淋巴管密度(LVD)和VEGF-C蛋白的表达。结果:VEGF-C mRNA和蛋白高表达于3株胃癌细胞,而VEGFR-3 mRNA和蛋白在上述3个细胞株中均呈弱表达。VEGF-C和VEGFR-3 mRNA均表达于3对伴有淋巴结转移的胃癌和相应的正常黏膜组织中,但在正常组织中的表达水平低于肿瘤组织。VEGF-C蛋白在66.3%(57/86)的病例中呈阳性表达。在伴淋巴结转移的胃癌中,VEGF-C表达较无淋巴结转移者更显著(P<0.001),其表达与淋巴管浸润(P<0.001)和TNM分期(P<0.01)均密切相关,但与病人的年龄和性别、肿瘤大小、位置、组织学类型、浸润深度及远处转移均无明显相关。LVD则与VEGF-C蛋白表达(P<0.001)、淋巴结转移(P<0.01)、淋巴管浸润(P<0.01)、TNM分期(P<0.05)及组织学类型(P<0.05)密切相关,但与病人的年龄和性别、肿瘤大小、位置、浸润深度及远处转移无明显相关。结论:在胃癌中,VEGF-C可能通过VEGFR-3信号通道促进淋巴管生成,从而增加胃癌淋巴结转移率。因此,肿瘤淋巴管生成可能成为治疗胃癌的一个新靶点。     

血管内皮生长因子-C在大肠癌中的表达及其与淋巴结转移的关系

目的研究血管内皮生长因子-C(VEGF-C)在大肠癌中的表达,探讨其与淋巴结转移的关系。方法应用免疫组织化学染色法检测94例大肠癌组织标本中VEGF-C的表达,同时应用逆转录-聚合酶链反应(RT-PCR)检测其mRNA在4株大肠癌细胞株中的表达。结果VEGF-C在53.2%的大肠癌患者中呈阳性表达;VEGF-C在淋巴结转移阳性组中的表达,与阴性组比较差异有统计学意义(P<0.01);VEGF-C的表达与淋巴管浸润和Dukes分期密切相关(P<0.01),但与年龄、性别、肿瘤的位置、浸润深度和血管浸润均无明显相关。VEGF-CmRNA表达于大肠癌LoVo及LoVo-5-Fu耐药细胞株。结论VEGF-C的表达可能参与肿瘤淋巴管生成,与大肠癌淋巴结转移密切相关。     

淋巴管生成因子与消化道肿瘤

目的 介绍VEGF-C和VEGF-D在淋巴管生成及消化道肿瘤淋巴转移中的作用的有关研究进展。方法对近年来VEGF-C和VEGF-D的结构功能特点、及其在淋巴管生成和消化道肿瘤淋巴转移中的作用的研究进展进行综述和分析。结果VEGF-C和VEGF-D可促进血管、淋巴管生成和淋巴转移。胃癌中VEGF-CmRNA及其蛋白表达与淋巴侵犯、静脉侵犯、淋巴结转移及低的5年生存率相关；VEGF-C在食管癌和结直肠癌中的作用以及VEGF-D在结直肠癌中的作用有待进一步研究。结论VEGF-C、VEGF-D／VEGFR-3信号通路可能成为控制肿瘤生长转移的理想靶点，抗新生淋巴管治疗有望成为肿瘤生物治疗新模式。     

Expression of vascular endothelial growth factor C (VEGF-C) and VEGF receptor-3 in human prostate cancer is associated with regional lymph node metastasis

BACKGROUND: Vascular endothelial growth factor C (VEGF-C) and its receptor, VEGFR-3, have been implicated as important factors in the formation of lymphatic vessels and recent evidence suggests that tumor lymphangiogenesis promotes lymphatic metastasis. METHODS: The expression of VEGF-C and VEGFR-3 was examined in 22 human prostate cancer specimens with immunohistochemistry. A semi-quantitative scoring system was used for evaluation of staining. RESULTS: Expression of VEGF-C was stronger in prostate cancer areas in comparison to adjacent benign glands. In addition, patients with lymph node metastases had a significantly higher expression of VEGF-C than patients without lymph node metastases. Interestingly, VEGFR-3 was expressed in malignant prostate epithelial cells and its expression was significantly higher in the lymph node positive group compared to the lymph node negative group. CONCLUSIONS: The results of the present study indicate that increased expression of VEGF-C and VEGFR-3 play a role in prostate cancer progression and in metastasis to regional lymph nodes.     

COX-2和VEGF-C表达与NSCLC淋巴转移的关系

目的探讨环氧化酶-2(COX-2)和血管内皮生长因子-C(VEGF-C)表达与淋巴管形成和淋巴结转移之间的关系。方法用免疫组化链霉素抗生物素蛋白-过氧化物酶(SP)法检测65例非小细胞肺癌(NSCLC)及16例正常肺组织中COX-2、VEGF-C及其受体VEGFR-3的表达;以VEGFR-3作标记计数肿瘤LVD,并结合临床病理特征进行统计学分析。结果65例NSCLC中COX-2和VEGF-C表达阳性率分别为76.9%(50/65)、72.3%(47/65)。COX-2表达与淋巴结转移(r=0.489,P<0.01)、临床分期(r=0.354,P<0.05)、VEGF0C(r=0.640,P<0.01)和LVD(r=0.518,P<0.01)呈正相关,而与病理分化程度呈负相关(r=-0.427,P<0.01)。VEGF-C表达与淋巴结转移(r=0.453,P<0.01)、临床分期(r=0.442,P<0.01)和LVD(r=0.624,P<0.01)呈正相关,与病理分化程度也呈负相关(r=-0.525,P<0.01)。结论NSCLC中COX-2与VEGF-C均高表达,COX-2可能通过VEGF-C促进肿瘤淋巴管生成和淋巴结转移。研究COX-2和VEGF-C在肿瘤中的协同作用有助于揭示肿瘤侵袭和转移的机制。     

血管内皮生长因子-C在胃癌中的表达及其与淋巴结转移的关系

目的　研究血管内皮生长因子 C(VEGF C)在胃癌中的表达并探讨其与淋巴结转移的关系。方法　应用逆转录 聚合酶链反应 (RT PCR)检测VEGF CmRNA在 5株胃癌细胞株中的表达情况 ,同时采用免疫组织化学法 ,检测 63例接受根治性切除手术病例的胃癌组织标本VEGF C蛋白的表达。结果　VEGF CmRNA表达于胃癌细胞株MKN 45、SGC 790 1及AGS。VEGF C蛋白则在 5 2 .4%(3 3 /63 )的病例中呈阳性表达。在伴淋巴结转移的胃癌中 ,VEGF C表达较无淋巴结转移者更显著 (P <0 .0 1)。同时 ,VEGF C表达与淋巴管浸润和TNM分期密切相关 (P <0 .0 1) ,但与年龄、性别、肿瘤大小、位置、Lauren分型、浸润深度和血管浸润均无明显相关。结论　VEGF C的表达与胃癌淋巴结转移密切相关。     

人结直肠癌VEGF-C的表达及与淋巴结转移之间的关系

目的 :探讨血管内皮生长因子C(VEGF C)在人结直肠癌中的表达及与肿瘤淋巴结转移之间的关系。方法 :选择 47例结直肠癌标本 ,应用免疫组织化学染色方法 ,检测VEGF C在人结直肠癌组织中的表达 ,同时对伴与不伴有淋巴结转移的结直肠癌VEGF C表达的差异进行统计学分析。结果 :VEGF C在结直肠癌组织中的阳性表达率为 44 .7%,其中有淋巴结转移的表达率为 6 0 .0 %,无淋巴结转移的表达率为 17.7%,二者差异有统计学意义 (P <0 .0 1)。结论 :结直肠癌可表达VEGF C ,并与肿瘤细胞淋巴转移有关。     

血管内皮生长因子-C和生存素在胃癌组织中的表达及其临床意义

目的 探讨胃癌组织中血管内皮生长因子（vascular endothelial growth factor，VEGF）-C和生存素（sunrivin）蛋白表达及其临床意义。方法 采用免疫组织化学SP法检测97例原发性胃癌组织、癌旁组织及20例正常胃黏膜组织中VEGF-C和survivin蛋白的表达，并分析其与临床病理特征和预后的关系。结果 胃癌组织VEGF-C和survivin蛋白表达阳性率分别为66．0％和57．2％，显著高于癌旁组织和正常胃组织（P〈0．05）；VEGF-C蛋白表达与肿瘤分化程度、肿瘤部位、肿瘤直径、静脉侵犯及远处转移等无关，但与淋巴结转移、淋巴管侵犯、浆膜面受累和肿瘤TNM分期等密切相关；survivin蛋白表达与肿瘤分化程度、肿瘤部位、肿瘤直径、静脉侵犯等无关，但与浆膜面受累、淋巴管侵犯、淋巴结转移、远处转移和肿瘤TNM分期等密切相关；VEGF-C和survivin阳性表达组术后生存率明显低于阴性组；VEGF-C和survivin在胃癌组织中阳性表达呈正相关。结论 VEGF-C及survivin蛋白的阳性表达可作为胃癌预后不良的参考指标。     

VEGF-C、VEGFR-3在NSCLC中的表达与淋巴转移

目的 探讨VEGF-C及VEGFR-3与NSCLC淋巴转移间的关系,为了解患者预后提供依据。方法 采用免疫组化法检测65例NSCLC中VEGF-C及VEGFR-3的表达,结合临床病理特征进行分析。结果 NSCLC中VEGF-C和VEGFR-3的阳性率明显高于正常肺组织(P0.05)。二者在Ⅲ期NSCLC中的表达率明显高于Ⅰ、Ⅱ期,且随病理分级的增高而显著增加(P0.05),在有淋巴结转移者明显高于无淋巴结转移者(P0.05)。结论 VEGF-C及VEGFR-3在NSCLC中高表达,与其临床分期、病理分级、病理类型和淋巴结转移均密切相关。     

前列腺癌组织中HIF-1α和VEGF-C及其受体VEGFR-3的表达及意义

目的探讨缺氧诱导因子-1α（hypoxia inducible factor-1,HIF-1α）、血管内皮生长因子-C（vascular endothelial growth factor-C,VEGF-C）和VEGF受体-3（vascular endothelial growth factor receptor-3,VEGFR-3）蛋白在前列腺癌组织中的表达及意义。方法应用免疫组织化学SP法测定64例前列腺癌和15例正常前列腺组织中HIF-1α、VEGF-C和VEGFR-3蛋白的表达情况。结果 HIF-1α、VEGF-C和VEGFR-3蛋白在前列腺癌组织中的表达率分别为60.9%、70.3%和71.9%,在正常前列腺组织中均无表达;HIF-1α蛋白的表达与病理分级、临床分期和淋巴转移有关,VEGF-C蛋白与病理分级和淋巴转移有关,VEGFR-3蛋白与淋巴转移有关（P〈0.05）;在前列腺癌组织中HIF-1α与VEGF-C、VEGFR-3;VEGF-C与VEGFR-3的表达呈正相关（P〈0.05）。结论前列腺癌组织中HIF-1α、VEGF-C和VEGFR-3蛋白的高表达与前列腺癌的发生及淋巴转移相关。     

淋巴管内皮生长因子-C在膀胱移行细胞癌中的表达及其临床意义

目的 :探讨具有促进淋巴管内皮细胞增殖和毛细淋巴管增生的淋巴管内皮生长因子 C(VEGF C)在膀胱移行细胞癌 (BTCC)中的表达及其与肿瘤淋巴管浸润、淋巴结转移等临床病理因素的关系。方法 :采用免疫组织化学方法研究 4 5例BTCC中VEGF C的表达水平 ,观察评估VEGF C的阳性表达与淋巴管浸润、淋巴结转移等临床病理资料的相关性。结果 :4 5例中 ,VEGF C阳性 36例 ,VEGF C的阳性表达与肿瘤分级、分期、静脉或淋巴管浸润、淋巴结及前列腺侵犯明显相关 (P <0 .0 5 ) ;多因素分析显示VEGF C的表达是唯一影响盆腔淋巴结转移的独立因素 (P =0 .0 0 3)。结论 :VEGF C与BTCC淋巴管浸润和转移有显著相关性 ,检测BTCC中VEGF C的表达能够预示盆腔淋巴结是否转移。     

淋巴管透明质酸受体1和血管内皮生长因子受体3在胃癌组织中的表达

目的以淋巴管透明质酸受体1（LYVE-1）和血管内皮生长因子受体3（VEGFR-3）作为淋巴管特异性标志物,研究胃癌组织和癌周组织中淋巴管的生成表达情况。方法应用免疫组织化学和Real—time聚合酶链反应的方法检测62例胃癌、癌周组织中的LYVE-1和VEGFR－3蛋白及其mRNA表达情况,计算淋巴管的密度（LVD）,检测淋巴管mRNA的相对含量。结果胃癌组织周边区淋巴管的密度高于胃癌组织中心区,而癌旁正常组织中很少见LYvE-1和VEGFR-3染色阳性的微淋巴管;胃癌组织周边区LVD和胃癌组织中LYVE-1和VEGFR-3 mRNA含量与胃癌组织浸润深度、淋巴结转移、肿瘤分期及术后复发有关（P〈0．05）。结论LYVE-1是一种特异性高于VEGFR-3的淋巴管内皮特异性标志物,淋巴管生成促进胃癌发展及淋巴结转移。     

血管内皮生长因子 C表达和淋巴管生成与结肠癌进展及预后的关系

目的评估血管内皮生长因子 C( VEGF- C)、淋巴管密度( LMVD)与结肠癌临床病理指标及预后的关系.方法用免疫组织化学法检测 44例原发结肠癌 VEGF- C和 VEGF受体- 3( VEGF R- 3)表达,计数 LMVD,分析上述指标与临床病理指标和预后的关系.结果本组结肠癌 VEGF- C阳性表达率为 43.2%( 19/44), LMVD为 10.14± 4.19. VEGF- C表达与肿瘤分化程度( P=0.003)、淋巴结转移( P=0.002)和 Dukes分期( P=0.001)相关. LMVD与淋巴结转移( P=0.001)和 Dukes分期( P=0.001)相关. VEGF- C表达阳性组 LMVD为 11.34± 4.83,高于 VEGF- C表达阴性组的 9.24± 3.48,但 VEGF- C与 LMVD无相关性( P=0.105). VEGF- C阳性组患者生存率明显低于阴性组( P=0.0225), LMVD阳性组患者生存率明显低于阴性组( P=0.0036).远处转移( P=0.0004)、淋巴结转移( P=0.021)和 LMVD( P=0.0469)可以作为结肠癌独立的预后因素.结论 VEGF- C和 LMVD对于判断结肠癌侵袭性和预后有重要参考价值. LMVD可以作为判断结肠癌预后的独立指标.     

miR-21和PDCD4mRNA在大肠癌组织中的表达及其临床意义

目的 探讨miR-21和PDCD4mRNA在大肠癌组织中的表达及与其临床病理特征的关系,并阐明在大肠癌活体组织中miR-21与其靶基因PDCD4表达的关系.方法 用荧光定量PCR技术检测43对大肠癌组织及其对应的正常黏膜组织中的miR-21和PDCD4 mRNA表达量;免疫组化SP法检测PDCD4蛋白的表达.结果 大肠癌组织中miR-21的表达升高(P＜0.05),而PDCD4mRNA的表达降低(P＜0.05).miR-21的表达与大肠癌的淋巴结转移、临床分期(Ⅰ+Ⅱ、Ⅲ+Ⅳ)有关(P＜0.05),而与患者的性别、肿瘤位置、肿瘤大小、浸润深度、远处转移、临床分期(Ⅰ、Ⅱ)无关(P＞0.05);PDCD4mRNA的表达与大肠癌的浸润深度、临床分期(Ⅰ、Ⅱ)有关(P＜0.05),而与患者的性别、肿瘤位置、肿瘤大小、淋巴结转移、远处转移、临床分期(Ⅰ+Ⅱ、Ⅲ+Ⅳ)无关(P＞0.05).miR-21的表达与PDCD4蛋白的核表达、核与浆表达之和存在负相关(P＜0.05),而与PDCD4mRNA、PDCD4蛋白的浆表达无相关性(P＞0.05).结论 miR-21和PDCD4mRNA在大肠癌中的异常表达与大肠癌的预后有关;在大肠癌活体组织中,miR-21通过与PDCD4mRNA的3’非编码区结合抑制PDCD4mRNA翻译成蛋白质.     

Overexpression of vascular endothelial growth factor-C correlates with lymph node micrometastasis in submucosal esophageal cancer

Lymph node metastasis, including lymph node micrometastasis (LMM), is one of the most important prognostic factors in esophageal squamous cell carcinoma (ESCC). Vascular endothelial growth factor C (VEGF-C) plays a key role in the process of lymphangiogenesis. We examined VEGF-C expression and tumor microvessel density of the primary tumors in ESCC and analyzed relationships between VEGF-C expression and clinicopathologic findings including LMM in submucosal ESCC. The subjects were 87 patients with submucosal ESCC. Immunohistochemical staining of VEGF-C and CD34 was performed with primary tumors, and staining of cytokeratin was performed with dissected lymph nodes. Microvessel density was calculated from CD34 expression, and LMM was detected by cytokeratin staining. VEGF-C overexpression significantly correlated with depth of tumor invasion, lymphatic invasion, and lymph node metastasis (P<0.05, P<0.0001, and P<0.0001, respectively). High microvessel density also correlated with lymphatic invasion and lymph node metastasis (P<0.005 and P<0.05, respectively). LMM was detected in 8 cases and 14 lymph nodes by cytokeratin staining. VEGF-C overexpression and high microvessel density were found in tumors with lymph node metastasis and/or LMM, compared with tumors without nodal metastasis or LMM (P<0.0001 and P<0.01, respectively). The present findings indicate that in ESCC with submucosal invasion, VEGF-C overexpression of the primary tumor is a strong high risk factor for lymph node metastasis, including LMM. Supported in Grant No. 17390373 part by grants-in-aid for scientific research from the Ministry of Education, Science, Sports and Culture, Japan. (Shoji Natsugoe, M.D., Ph.D.)     

Vascular Endothelial Growth Factor C Promotes Lymph Node Metastasis in a Rectal Cancer Orthotopic Model

Purpose Vascular endothelial growth factor C (VEGF-C), a novel member of the vascular endothelial growth factor family, is a relatively specific lymphangiogenic growth factor. It has been suggested that increased expression of VEGF-C in primary tumors is correlated with lymph node metastasis. We conducted this study to determine whether VEGF-C directly affects lymphangiogenesis and lymph node metastasis in colorectal cancer.Methods For an accurate analysis and clear visualization of metastases, the rectal cancer cell line, DLD1, was engineered to stably express green fluorescent protein (GFP) (DLD1/GFP). We implanted DLD1/GFP cells overexpressing VEGF-C orthotopically into the rectal walls of nude mice.Results Lymph node metastasis was confirmed in all (100%) of the mice bearing DLD1/GFP-VEGF-C tumors, but in only 25% of the mice bearing control tumors. There were more lymph node metastases per mouse in the mice bearing DLD1/GFP-VEGF-C tumors than in the mice bearing control tumors. There were no differences in cell growth and motility in vitro or in the resulting tumor volume from the implanted cells between the two groups. Immunohistochemical staining revealed that VEGF-C induced the growth of lymphatic vessels, which were enlarged in the tumor periphery and contained tumor cell emboli.Conclusion These results suggest that VEGF-C-induced lymphangiogenesis mediates tumor spread and the formation of lymph node metastasis.     

大肠癌中VEGF-C分泌水平的检测及其意义

目的 检测大肠癌患者血清及不同的细胞系中血管内皮生长因子C（VEGF-C）和血管生长因子（VEGF）的分泌水平，评估其在诊断大肠癌淋巴结转移中的意义。方法 采用酶联免疫法检测66例大肠癌（观察组）和30例健康者（对照组）血清中及4株大肠癌细胞系培养液中VEGF-C和VEGF的分泌的水平。结果 观察组血清VEGF-C和VEGF均高于对照组（P〈0．01）；有淋巴结转移组的血清VEGF-C和VEGF均高于无淋巴结转移组（P〈0．01）；有淋巴管和／或血管浸润组上述两指标均高于无浸润组（P〈0．01）。血清VEGF-C水平在1438．0pg／mL以上者其灵敏度为81．0％。特异性为76．0％；血清VEGF水平在240．2pg／mL以上者其灵敏度为72．0％，特异性为74．0％。联合检测VEGF-C和VEGF对淋巴结转移阳性率的预测价值达84．6％，阴性预测价值达94．4％，精确度达93．7％。LoVo及LOVo-5FU培养液中VEGF-C分泌水平明显升高（P〈0．01）。结论 检测VEGF-C为判断大肠癌患者淋巴结是否转移可能提供新的诊断依据。VEGF-C可望成为治疗大肠癌淋巴道转移的新靶点。     

VEGF-C与VEGF-D蛋白的表达对结直肠癌淋巴结转移的影响

目的探讨VEGF-C与VEGF-D蛋白在结直肠癌组织中的表达,及其与结直肠癌淋巴转移的关系。方法采用Western blot方法检测80例结直肠癌组织中VEGF-C与VEGF-D的表达水平。结果在结直肠癌组织中,有淋巴结转移组比无淋巴结转移组VEGF-C、VEGF-D两种蛋白的表达水平高（P〈0.05）;其中在有淋巴结转移的结直肠癌组织中,N2期比N1期VEGF-C、VEGF-D两种蛋白的表达水平高（P〈0.05）。结论 VEGF-C与VEGF-D蛋白的表达与结直肠癌淋巴结转移的发生发展有关,用Western blot方法检测两者的表达水平对判断结直肠癌生物学行为和预后具有参考价值。     

Expression of vascular endothelial growth factor receptor-3 (VEGFR-3) in human prostate

BACKGROUND: The growth and dissemination of tumors has been associated with angiogenesis, which is regulated by a group of polypeptide factors including vascular endothelial growth factor-C (VEGF-C). VEGF-C binds its receptor, vascular endothelial growth factor receptor-3 (VEGFR-3) to promote growth of tumor-associated lymphatic vessels. METHODS: In this study, microarray technology was used to build tissue arrays of normal prostate, benign prostate hyperplasia (BPH) and prostate carcinomas (PCa) using tissues from 640 patients. Slides were sectioned and stained with a polyclonal antibody to VEGFR-3 using a standard immunoperoxidase method and digitized. Immunoreactivity was scored using a 0-3+ semiquantitation scoring system for both intensity and percentage. The sum index was obtained by totaling the scores. RESULTS: VEGFR-3 is expressed in normal prostate, BPH, and PCa, but VEGFR-3 expression is up-regulated in PCa. We also found that VEGFR-3 is correlated with pre-operative prostate-specific antigen (Pre-PSA), Gleason score, and lymph node metastasis. The recurrence-free 5-year survival in cases with lower sum index (0-3) was significantly higher than that in cases with higher sum index (4-6) (77.3, 69.6%, respectively, P = 0.037) by Kaplan-Meier actuarial model. CONCLUSIONS: Our data suggest that VEGFR-3 expression is associated with tumor progression and may play an important role in facilitating lymphatic spread of PCa; high-level of VEGFR-3 expression in prostate cancer cells increases the risk of biochemical recurrence in prostate cancer patients treated by radical prostatectomy.     

胃癌组织中血管内皮生长因子C的表达与淋巴结微转移的关系

目的：检测胃癌患者血管内皮因子C（VEGF-C）蛋白及mRNA在胃癌组织中的表达情况,探讨其与胃癌淋巴结微转移的关系。方法应用免疫组织化学法及RT-PCR法检测80例胃癌组织、癌旁组织及正常组织中的VEGF-C蛋白及mRNA表达情况;免疫组化法检测淋巴结微转移情况,比较有无淋巴结微转移的VEGF-C蛋白和mRNA的表达差异,明确存在的关联性。结果VEGF-C蛋白及mRNA两者在胃癌组织中明显高于癌旁组织和正常组织（P＜0.05）。结论胃癌组织中的VEGF-C蛋白及mRNA与胃癌淋巴结微转移相关,基因检测优于蛋白,可作为评价胃癌患者是否存在淋巴结微转移的优选指标。