The balance of VEGF-C and VEGFR-3 mRNA is a predictor of lymph node metastasis in non-small cell lung cancer

A positive association between vascular endothelial growth factor-C (VEGF-C) expression and lymph node metastasis has been reported in several cancers. However, the relationship of VEGF-C and lymph node metastasis in some cancers, including non-small cell lung cancer (NSCLC), is controversial. We evaluated the VEGF-C and vascular endothelial growth factor receptor-3 (VEGFR-3) expression in NSCLC samples from patients who had undergone surgery between 1998 and 2002 using real-time quantitative RT-PCR and immunohistochemical staining. We failed to find a positive association between VEGF-C and VEGFR-3 mRNA expression and lymph node metastasis in NSCLC. An immunohistological study demonstrated that VEGF-C was expressed not only in cancer cells, but also in macrophages in NSCLC, and that VEGFR-3 was expressed in cancer cells, macrophages, type II pneumocytes and lymph vessels. The VEGF-C/VEGFR-3 ratio of the node-positive group was significantly higher than that of the node-negative group. Immunohistochemical staining showed that VEGFR-3 was mainly expressed in cancer cells. The immunoreactivity of VEGF-C and VEGFR-3 was roughly correlated to the mRNA levels of VEGF-C and VEGFR-3 in real-time PCR. VEGF-C mRNA alone has no positive association with lymph node metastasis in NSCLC. The VEGF-C/VEGFR-3 ratio was positively associated with lymph node metastasis in NSCLC. This suggests that VEGF-C promotes lymph node metastasis while being influenced by the strength of the VEGF-C autocrine loop, and the VEGF-C/VEGFR-3 ratio can be a useful predictor of lymph node metastasis in NSCLC.     

Deguelin—An inhibitor to tumor lymphangiogenesis and lymphatic metastasis by downregulation of vascular endothelial cell growth factor‐D in lung tumor model

Deguelin, a rotenoid of the flavonoid family, has been reported to possess antiproliferative and anticarcinogenic activities in several cell lines and tumor models. However, it is still unclear whether deguelin effectively inhibits tumor‐associated lymphangiogenesis and lymphatic metastasis. Since tumor production of vascular endothelial cell growth factor (VEGF)‐D was associated with tumor lymphangiogenesis and lymphatic metastasis, we established the mouse lymphatic metastasis model by transfecting high expression VEGF‐D into LL/2 Lewis lung cells (VEGF‐D‐LL/2) and explored the effects of deguelin on lymphatic metastasis in the immunocompetent C57BL/6 mice. Our results indicated that deguelin inhibited proliferation, migration of VEGF‐D‐LL/2 cells via downregulating AKT and mitogen‐activated protein kinase pathway and interfered tube formation of lymphatic vascular endothelial cells on matrigel at nanomolar concentrations. Deguelin significantly downregulated the expression of VEGF‐D both at mRNA and protein levels in VEGF‐D‐LL/2 cells in a dose‐dependent manner. In the in vivo study, intraperitoneal administration of deguelin (4 mg/kg) remarkably inhibited the tumor‐associated lymphangiogenesis and lymphatic metastasis. The rates of lymph node and lung metastasis in deguelin‐treated mice were 0 and 16.7% compared with 58.3 and 83.3% in control group mice, respectively. Deguelin also resulted in a remarkable delay of tumor growth and prolongation of life span. Immunohistochemical staining with antibodies against VEGF‐D, LYVE‐1 and VEGFR‐3 revealed fewer positive vessel‐like structures in deguelin‐treated mice compared with control group mice. Taken together, we demonstrate for the first time that deguelin suppresses tumor‐associated lymphangiogenesis and lymphatic metastasis by downregulation of VEGF‐D both in vitro and in vivo.     

VEGF-C及其受体VEGFR3在乳腺癌的表达、定位及其在淋巴结转移中的作用

目的:研究VEGF-C及其VEGFR3在乳腺癌组织及癌周组织中的表达情况、分布定位及其在乳腺癌淋巴结转移中的作用。方法:采用免疫组化SP法检测70例患者乳腺癌组织及其癌周组织中VEGF-C和VEGFR3的表达情况,CK19染色检测腋窝淋巴结微转移情况。结果:乳腺癌70例,VEGF-C在乳腺癌组织阳性表达率为78.6%,在癌周组织中的表达率为54.3%(P<0.01)。VEGFR3阳性的脉管数在乳腺癌组织为7.90±5.57,瘤周组织中为10.31±6.52(P<0.01)。VEGFR3的表达主要受乳腺癌细胞VEGF-C表达的影响,两者具有明显的相关性(r=0.66,P<0.05)。乳腺癌淋巴管主要定位于癌周组织及肿瘤间质内,癌巢内未发现成熟淋巴管。VEGF-C在乳腺癌组织内的阳性表达率淋巴结微转移阳性组为87.5%,在淋巴转移阴性组为71.1%,两组差异有统计学意义(P<0.01)。VEGFR3阳性脉管数在乳腺癌瘤周组织中淋巴转移阳性组为12.48±6.51,淋巴转移阴性组为8.47±6.02,两组差异有统计学意义(P<0.01)。结论:VEGF-C在乳腺癌组织肿瘤细胞中呈高表达,而其受体VEGFR3则表达乳腺癌间质和癌周组织的淋巴内皮细胞;乳腺癌新生淋巴管主要定位于癌周及间质;VEGF-C在乳腺癌组织、VEGFR3在癌周组织的表达与乳腺癌淋巴浸润和淋巴结转移密切相关。     

人参皂苷免疫纳米对人胃癌裸鼠移植瘤生长抑制机制探讨

目的研究人参皂苷免疫纳米（VEGFR3-mediated immune-nanoemulsion of ginsenoside Rg3,VRIN）对胃癌生长及其淋巴管生成的机制。方法通过外科原位移植能表达红色荧光蛋白人胃癌NUGC-4细胞的方法建立裸鼠胃癌模型,将32只Balb/c无胸腺裸鼠随机分为生理盐水组、5-氟尿嘧啶（5-FU）组（20mg/kg）、VRIN高剂量组（1mg/kg）和VRIN低剂量组（0.1mg/kg）4组,每组8只。实验终点处死所有受试裸鼠,在开放荧光系统下观察肿瘤生长及转移情况,切除移植肿瘤测瘤质量和瘤体积。免疫组织化学法检测肿瘤组织中微淋巴管密度（lymphatic microvessel density,LMVD）,Real time-PCR和免疫组织化学法检测VEGF-C蛋白和mRNA表达。结果实验终点开放体内荧光成像显示,胃癌NUGC-4细胞肿瘤生长抑制率VRIN高剂量为72.9%,低剂量组为14.5%,5-FU组为69.2%,VRIN高剂量组和5-FU组较生理盐水组瘤质量明显减轻,P〈0.001。生理盐水组淋巴转移率为87.5%（7/8）,5-FU组为50.0%（4/8）,VRIN高剂量组为12.5%（1/8）,低剂量组为37.5%（3/8）,VRIN高剂量组与生理盐水组差异有统计学意义,P=0.01。LMVD计数结果显示,生理盐水组为9.29±2.06,5-FU组为6.42±1.38,VRIN高剂量组为4.25±1.08,差异有统计学意义,F=20.895,P〈0.001;VRIN高剂量组VEGF-C蛋白表达量为0.190±0.059,与生理盐水组（0.269±0.051）差异有统计学意义,P=0.014;VEGF-C mRNA表达结果显示,生理盐水组为1.05±0.19,5-FU组为0.62±0.25,VRIN高剂量组为0.49±0.19,差异有统计学意义,F=8.190,P=0.002。结论 VRIN可抑制人胃癌细胞祼鼠移植瘤生长及淋巴结转移,并通过下调VEGF-C表达抑制肿瘤淋巴管生成。     

淋巴管生成因子在肿瘤转移中的作用

肿瘤转移是恶性肿瘤高死亡率的主要原因,淋巴管系统是肿瘤转移的一个重要途径。长期以来,由于缺乏淋巴管内皮细胞特异性标志,我们对淋巴管系统及其在肿瘤转移中的作用知之甚少。目前,淋巴管生成因子和淋巴管内皮细胞特异性标志的相继发现大大促进了淋巴系统这一领域的研究。大量动物实验以及临床病理研究结果显示,淋巴管生成因子与肿瘤中淋巴管生成和淋巴结转移显著相关,这使得淋巴管生成因子成为肿瘤淋巴道转移机制和抗肿瘤研究的新热点。     

VEGF-C与肿瘤淋巴管生成和淋巴结转移关系研究进展

淋巴管生成是实体肿瘤发生淋巴结转移的重要步骤,但与肿瘤血管生成相比,目前对淋巴管生成的研究较少。随着研究的不断深入,在肿瘤的动物模型以及人类肿瘤组织和肿瘤周边组织中,均发现了新生淋巴管。目前研究已证实,血管内皮生长因子-C(VEGF-C)在肿瘤的淋巴管生成及淋巴结转移中发挥重要作用,为抗肿瘤淋巴转移治疗提供了有效的靶点,有助于改善患者预后、延长生存期。     

VEGF-C和VEGFR-3对乳腺癌淋巴管生成和淋巴结转移的影响

目的:探讨VEGF-C及VEGFR-3对乳腺癌淋巴结转移的影响。方法:采用免疫组化SP法检测乳腺病、乳腺纤维腺瘤和乳腺癌中VEGF-C及VEGFR-3的表达状态。结果:10例乳腺病及纤维腺瘤VEGF-C均阴性,也无VEGFR-3阳性淋巴管;48例乳腺癌组织VEGF-C阳性率为70.83%(34/48);淋巴结转移者VEGF-C阳性率为92.59%(25/27),显著高于无转移者(42.86%,9/21)(P＜0.01)。VEGFR-3阳性淋巴管条数随VEGF-C强度增强而增多。在淋巴结转移者中,VEGFR-3阳性淋巴管条数(6.03)多于无淋巴结转移者(2.01)(P＜0.01)。结论:乳腺癌组织中VEGF-C的表达状态与VEGFR-3阳性淋巴管数量及淋巴结转移关系密切。     

Neuropilin-1与肿瘤血管新生和生长转移

神经纤毛蛋白-1（Neuropilin-1）最早是作为轴突导向分子collapsin／semaphorin的受体被发现，在神经发育过程中引导轴突选择正确途径以成功到达靶区，与此同时又可作为血管内皮生长因子受体-2（VEGFR-2）的共受体表达于血管内皮细胞，在血管新生中发挥作用。近来的研究表明NRP-1可通过依赖于VEGF和独立于VEGF的方式参与调节肿瘤血管新生，在肿瘤生长转移中发挥作用。     

Effects of VEGFR-3 phosphorylation inhibitor on lymph node metastasis in an orthotopic diffuse-type gastric carcinoma model

Background:

Vascular endothelial growth factor receptor-3 (VEGFR-3) signalling mediates lymphangiogenesis and lymphatic invasion; however, the effect of VEGFR-3 inhibition on the lymph node (LN) metastasis remains unclear. The aim of this study is to clarify the benefit of a VEGFR-3 inhibitor Ki23057 for LN metastasis.

Methods:

Ki23057 was administered orally to gastric cancer models created by orthotopic inoculation of diffuse-type gastric cancer cells, OCUM-2MLN. The effects of Ki23057 on lymphatic vessel invasion, lymphatic vessel density, and VEGFR-3 phosphorylation were examined by immunostaining or immunoblotting.

Results:

Ki23057 inhibited the autophosphorylation of VEGFR-3, with IC₅₀ values of 4.3 nM in the cell-free kinase assay. Murine gastric cancer models created by the orthotopic inoculation of OCUM-2MLN cells showed the diffusely infiltrating growth and frequently developed LN metastasis. The oral administration of Ki23057 significantly (P<0.01) reduced the size of orthotopic tumours and the number of the metastatic LN in gastric cancer models. The degree of lymphatic invasion and lymphangiogenesis was significantly (P<0.05) lower in the gastric tumours treated by Ki23057. Ki23057 inhibited the phosphorylation of VEGFR-3 of lymphatic endothelial cells in gastric tumours.

Conclusion:

The inhibition of lymphangiogenesis targeting VEGFR-3 phosphorylation is a therapeutic strategy for inhibiting LN metastasis of diffuse-type gastric cancer.     

子宫颈鳞状细胞癌VEGF-C及其受体 VEGFR-3的表达及其临床意义

目的 研究子宫颈鳞癌组织中血管内皮生长因子C(VEGF-C)及其受体VEGFR-3的表达和临床意义.方法 采用免疫组化方法 对10例正常子宫颈组织、12例CIN Ⅰ、12例CIN Ⅱ、20例CIN Ⅲ和66例子宫颈鳞癌的VEGF-C及其受体VEGFR-3的表达进行分析.结果 在子宫颈浸润性鳞癌中VEGF-C表达阳性率为75.8%,而正常子宫颈上皮、CIN Ⅰ、CIN Ⅱ和CIN Ⅲ则分别为30.0%、41.7%、41.7%和45.0%.浸润性鳞癌和正常上皮组比较差异具有显著性(P<0.01).鳞癌组织VEGF-C表达水平和间质VEGFR-3阳性淋巴管均数存在正相关性.子宫颈鳞癌VEGF-C表达水平和间质VEGFR-3阳性淋巴管数分别与FIGO分期和局部淋巴结转移呈正相关,差异均具有显著性.结论 VEGF-C表达增高和淋巴管增生在子宫颈鳞癌发生、发展过程中发挥促进作用,VEGF-C和VEGFR-3可作为子宫颈鳞癌组织的侵袭力和淋巴结转移预测的重要指标.     

血管内皮生长因子D及其受体3在乳腺癌淋巴转移中作用的研究进展

乳腺癌是女性最常见的恶性肿瘤。乳腺癌淋巴转移与患者的预后密切相关。最近研究表明淋巴管生成可能会主动促进淋巴转移的发生,而血管内皮生长因子家族的部分成员在这一过程中发挥了重要作用,如血管内皮生长因子C、血管内皮生长因子D及其受体3等。但是,对乳腺癌中血管内皮生长因子D的作用及其预后价值、血管内皮生长因子受体3与乳腺癌淋巴管生成的关系等问题仍有争议。本文对近年国内外有关血管内皮生长因子D及其受体3在乳腺癌淋巴转移中作用的研究进展作一综述。     

肿瘤淋巴管生成的研究进展

淋巴管对实体瘤的转移起着重要作用，目前研究表明，淋巴管生成因子（VEGF-C和VEGF-D）可通过激活其位于淋巴管内皮细胞上的受体（VEGFR-3）促进肿瘤内淋巴管生成，进而增加淋巴结转移率。同时，阻断VEGFR-3信号通道可抑制肿瘤淋巴管生成及淋巴结转移。因此，抗肿瘤淋巴管生成有望成为肿瘤转移抑制治疗的一条新途径。     

直肠癌组织PI3K和Survivin及VEGF表达与肿瘤侵袭转移相关性的探讨

目的:探讨磷脂酰肌醇3-激酶(PI3K)、存活素(Survivin)及血管内皮生长因子(VEGF)蛋白在人直肠癌组织中的表达,及其与直肠癌临床分期、组织学分级和侵袭转移的相关性及意义.方法:免疫组化SP法检测PI3K、Survivin和VEGF蛋白在60例直肠癌、30例直肠腺瘤及10例正常直肠组织中的表达.结果:PI3K在直肠癌、腺瘤和正常组织中的异常表达率分别为81.7％(49/60)、36.7％(11/30)和30.0％(3/10),差异有统计学意义,x2=17.86,P=0.001;Survivin的阳性表达率分别为90.0％(54/60)、80.0％(24/30)和20.0％(2/10),差异有统计学意义,x2=9.54,P=0.002;VEGF的异常表达率分别为71.7％(43/60)、43.3％(13/30)和30.0％(3/10),差异有统计学意义,x2=5.68,P=0.025.PI3K、Survivin及VEGF的异常表达均与临床分期及转移密切相关,P值均＜0.05.VEGF在直肠癌组织中染色阳性者(11.04±1.68)微血管密度(MVD)较染色阴性者(7.66±1.22)高,t=7.53,P＜0.05.在直肠癌组织中PI3K与Survivin(r=0.396,P=0.001)和VEGF (r=0.417,P=0.000 5)表达呈正相关,Survivin与VEGF表达呈正相关,r=0.332,P=0.005.PI3K和Survivin及VEGF蛋白协同阳性表达的侵袭转移率为55.0％(33/60),其中Ⅳ期远处转移率为93.8％(15/16).结论:PI3K和Survivin及VEGF蛋白的表达与临床分期及肿瘤侵袭转移密切相关,提示PI3K和Survivin及VEGF信号转导通路在人直肠癌侵袭转移中起重要作用.     

CXCL12/CXCR4轴在肿瘤生长和转移中的作用及其机制

近年来,趋化因子在肿瘤生长和转移中的作用引起人们的强烈关注。在多种肿瘤中（包括乳腺癌、胰腺癌、前列腺癌、结直肠癌等）发现,CXCL12/CXCR4分子对能够促进肿瘤细胞的生长,抑制肿瘤细胞的凋亡,促进肿瘤血管生成,影响某些肿瘤的靶向转移,增强肿瘤细胞的黏附和迁移能力,影响肿瘤细胞的分泌行为,提示该轴可能成为抗肿瘤药物的新靶点,具有潜在的临床应用前景。     

Association between intratumoral free and total VEGF, soluble VEGFR-1, VEGFR-2 and prognosis in breast cancer

Vascular endothelial growth factor (VEGF) receptors consist of three cell-membrane type receptors (VEGFR-1, VEGFR-2 and VEGFR-3), and soluble form of VEGFR-1 (sVEGFR-1), an intrinsic negative counterpart of the VEGF. In this study, we measured intratumoral protein levels of free and total VEGF, VEGFR-2 and sVEGFR-1 from 202 primary breast cancer tissues and examined their prognostic values. A significant inverse correlation was found between free or total VEGF and oestrogen receptor (ER) status (P=0.042 and 0.032, respectively). A univariate analysis showed that low sVEGFR-1 and high total VEGF were significantly associated with poor prognosis in disease-free survival (DFS) and overall survival (OS). The ratio of sVEGFR-1 to total VEGF was a strong prognostic indicator (DFS: P=0.008; OS: P=0.0002). A multivariate analysis confirmed the independent prognostic values of total VEGF and the ratio of sVEGFR-1 to total VEGF. In subgroup analysis, total VEGF was a significant prognostic indicator for ER-positive tumours but not for ER-negative tumours, whereas sVEGFR-1 was significant for ER-negative tumours but not for ER-positive tumours. In conclusion, the intratumoral sVEGFR-1 level, VEGF level and the ratio of sVEGFR-1 to total VEGF are potent prognostic indicators of primary breast cancer, and might be relevant to ER status.     

VEGF-C 及VEGFR-3 在肝细胞癌中的表达

 目的　探讨VEGF-C、VEGFR-3、微血管密度（MVD）、微淋巴管密度（LVD）及肝细胞癌临床病理特征之间的关系。方法　取肝细胞癌组织标本60例，正常肝组织标本20例。采用反转录聚合酶链反应（RT-PCR）方法分析其中VEGF-C及VEGFR-3 mRNA 的表达，以免疫组织化学法检测肝癌MVD及LVD，并分析四者与肝癌临床病理特点之间的关系。结果　肝癌组织VEGF-C、VEGFR-3 mRNA表达、MVD及LVD高于正常肝组织（P＜0.01）；肝癌组织中，VEGF-C与VEGFR-3表达、MVD及LVD均呈正相关（P＜0.01），VEGF-C及VEGFR-3表达与肝癌肝内转移、门静脉癌栓形成及淋巴转移相关（P＜0.01），MVD与肝癌肝内转移、门静脉癌栓形成相关（P＜0.01），LVD与淋巴转移相关（P＜0.01）。结论　肝细胞癌组织中VEGF-C及VEGFR-3表达增多，可能通过参与血管、淋巴管生成促进肿瘤的侵袭、转移。     

VEGF-D和VEGFR-3表达与食管鳞癌淋巴结转移相关性的探讨

目的:探讨VEGF-D/VEGFR-3信号通路与食管鳞癌淋巴结转移的关系。方法:采用逆转录多聚酶链反应技术(RT-PCR)检测VEGF-D及其受体VEGFR-3在52例食管鳞癌组织和20例正常食管组织中的表达,并分析其与食管鳞癌临床病理指标的关系。结果:在正常食管组织中VEGF-D和VEGFR-3mRNA表达均呈阴性;在食管鳞癌组织中,VEGF-DmRNA阳性表达32例(61.5%),VEGFR-3mRNA阳性表达28例(53.8%)。VEGF-DmRNA和VEGFR-3mRNA表达与食管鳞癌淋巴结转移显著相关,χ2值分别为15.333和15.594,P均<0.01;与浸润深度亦显著相关,χ2值分别为25.646和10.511,P均<0.05;VEGF-D与VEGFR-3表达呈正相关,χ2=25.142,P=0.001。结论:VEGF-D及VEGFR-3信号通路与食管鳞癌淋巴结转移及进展程度关系密切。     

淋巴管生长因子对食管鳞癌血管生成、肿瘤转移及预后的影响

目的：探讨淋巴管生长因子与食管鳞癌临床病理参数、血管生成及预后的关系。方法：采用Elivision二步免疫组化法检测51例食管鳞癌手术切除标本中VEGF—C的表达，应用CD34标记肿瘤内微血管，计数微血管密度，对VEGF—C与临床病理参数、肿瘤内MVD以及预后间的关系进行回顾性分析。结果：食管鳞癌淋巴结转移组VEGF—C阳性率和MVD明显增高；VEGF—C阳性组和高MVD组患者的中位生存时间以及3年、5年生存率均明显低于阴性组。VEGF—C与MVD呈正相关。结论：淋巴管生长因子VEGF—C可促进肿瘤微血管生成。VEGF—C阳性、MVD增高可作为食管鳞癌淋巴转移增加以及预后差的指标。     

VEGF-D/VEGFR-3信号通路在声门上型喉癌淋巴转移过程中的作用

目的:探讨VEGFD/VEGFR3信号通路在声门上型喉癌淋巴结转移过程中所起的调控作用。方法:应用RTPCR方法检测VEGFD及其受体VEGFR3在声门上型喉癌交界区癌组织、癌旁组织及正常喉黏膜组织的表达水平。结果:有淋巴转移组VEGFD及其受体VEGFR3癌组织表达分别为2.070±0.074和2.022±0.061,癌旁组织分别为1.129±0.067和1.131±0.068,正常喉黏膜组织分别为0.450±0.048和0.478±0.064;无淋巴转移组VEGFD及其受体VEGFR3癌组织表达分别为1.327±0.060和1.328±0.054,癌旁组织分别为0.927±0.052和0.917±0.061,正常喉黏膜分别为0.385±0.033和0.335±0.040;两组VEGFD及其受体VEGFR3表达均为癌组织高于癌旁组织高于正常喉黏膜组织,差异有统计学意义,t≥5.651,P<0.001。有淋巴转移组癌、癌旁组织二因子表达均高于无转移组,差异有统计学意义,t≥2.388,P<0.05;两组正常组织二因子表达差异无统计学意义,t≤1.873,P>0.05。结论:VEGFD/VEGFR3信号通路与声门上型喉癌淋巴结转移密切相关,二者在癌组织、癌旁组织的高表达有可能作为预测淋巴结转移的指标。VEGFD/VEGFR3通路将有望成为声门上型喉癌淋巴结转移预防和治疗的新靶点。