胃癌淋巴结转移与VEGF—D、VEGFR-3、LVD的关系

目的探讨胃癌淋巴结转移与血管内皮生长因子D（VEGF—D）、血管内皮生长因子受体3（VEGFR-3）、淋巴管密度（LVD）表达间的关系。方法对59例胃癌术后标本采用免疫组织化学sP法检测VEGF—D表达,用VEGFR-3抗体免疫组织化学sP法标记淋巴管,检测肿瘤组织及正常组织LVD。结果VEGF—D、VEGFR-3在癌组织中表达率分别为59．32％、67．80％,明显高于癌旁不典型增生组织及正常胃黏膜组织（P〈0．05）。癌周组织的LVD表达为（21．29±8．21）,明显高于周边正常组织（P〈0．05）。VEGF—D表达阳性组35例与阴性组24例的LVD分别为（23．15±7．58）和（11．93±5．31）,其差异具有统计学意义（P〈0．05）。VEGF—D、LVD在癌组织中的表达与淋巴结直径、淋巴结分期、浸润深度、TNM分期及分化程度有关（P〈0．05）。结论VEGF．D在胃癌组织中高表达,并与肿瘤的淋巴管形成及淋巴结转移密切相关。     

血管内皮生长因子-C,-D及其受体3在大肠癌淋巴转移和预后中的价值

目的探讨血管内皮生长因子(VEGF)-C,VEGF-D及其受体3(VEGFR-3)在大肠癌淋巴转移和预后中的价值。方法应用SABC免疫组化和RT-PCR方法检测80例大肠癌和30例大肠正常组织的VEGF-C、VEGF-D、VEGFR-3表达。随访、记录患者的临床病理参数和生存资料,分析其相关性。结果①大肠癌VEGF-C、VEGF-D和VEGFR-3均被染成棕黄褐色,三者蛋白表达阳性率分别为48．8%、56．3%、38．8%,相对表达量为1．09±1．20、1．13±1．09、0．90±1．19；正常大肠组织均无表达。与VEGF-C、VEGF-D和VEGFR-3的mRNA表达一致。②VEGF-C与VEGFR-3表达(P= 0．0069),VEGF-D与VEGFR-3表达间(P=0．0024)存在相关性；VEGF-C与VEGF-D无关。③VEGF-C、VEGF-D、VEGFR-3表达与大肠癌患者的年龄性别、肿瘤部位大小、大体分型、组织学分类、分化程度、肝、肺转移无关,但与Dukes分期(P=0．0234,P=0．0003,P=0．0429)、淋巴结转移(P= 0．0059,P＜0．01,P=0．0068)显著相关；且VEGF-C、VEGFR-3阳性大肠癌患者死亡率明显高于阴性患者(P=0．0374,P=0．0127),生存期明显短于阴性患者(P＜0．01,P＜0．01),VEGF-D则与预后无关。结论大肠癌细胞可分泌淋巴管生成因子VEGF-C、VEGF-D及其受体VEGFR-3,后者通过VEGF-C、-D/VEGFR-3信号通路诱导淋巴管内皮细胞新生和淋巴管生成,从而促进淋巴结转移和肿瘤生长；VEGF-C、VEGF-D、VEGFR-3可作为大肠癌的淋巴管生成的分子表型和淋巴转移及预后判断的重要指标。     

鼻咽癌VEGF-C、VEGFR-3表达及淋巴管密度与淋巴结转移、复发及预后的关系

采用免疫组化法检测鼻咽癌(NPC)患者活检组织标本中的血管内皮生长因子-C(VEGF-C)、血管内皮生长因子受体-3(VEGFR-3)表达及淋巴管密度(LVD),统计3a生存率,分析VEGF-C、VEGFR-3及LVD与淋巴结复发、转移及预后的关系.结果MPC有淋巴结转移者的VEGF-C、VEGFR-3阳性率及LVD均低于无淋巴结转移者(P均<0.01).单因素分析发现N分期及LVD与淋巴结复发有关,多因素分析发现仅N分期是影响NPc淋巴结复发的独立风险因素;高LVD及VEGF-C、VEGFR-3高表达者的3a生存率明显低于低表达者(P均<0.05).多因素分析表明,VEGF-C、LVD及临床分期是影响NPC患者生存率的独立因素.提示NPC高LVD患者可能增加淋巴结复发的风险;VEGF-C、VECFR-3高表达及高LVD患者预后较差.     

血管内皮生长因子及其受体在子宫内膜癌中的表达及对淋巴管生成的作用

目的检测子宫内膜癌中血管内皮生长因子(VEGF)-D及其受体VEGFR-3表达,探讨其在子宫内膜癌淋巴管新生中的价值。方法选择120例子宫内膜癌存档的蜡块作为观察组,60例正常增生期子宫内膜蜡块作为对照组,应用免疫组织化学技术检测VEGF-D、VEGFR-3及淋巴管内皮透明质酸受体(LYVE)-1的表达,探讨其临床意义及在淋巴管生成中的价值。结果 VEGF-D、VEGFR-3在子宫内膜癌中表达的阳性率明显高于对照组,LYVE-1标记的淋巴管密度明显高于对照组(P0.05),VEGF-D、VEGFR-3及LMVD与子宫内膜癌的浸润深度及淋巴结转移密切相关,肿瘤中VEGF-D与VEGFR-3、VEGF-D与LMVD、VEGFR-3与LMVD均呈正相关。结论子宫内膜癌中VEGF-D及VEGFR-3高表达,并具有协同作用,促进LYVE-1阳性淋巴管生成,联合检测VEGF-D、VEGFR-3及LYVE-1可能有助于判断子宫内膜癌的预后。     

大肠癌组织中生长抑素和血管内皮生长因子-C的表达及其临床意义

目的探讨大肠癌组织中生长抑素（SS）和血管内皮生长因子-C（VEGF-C）的表达及其临床意义。方法采用免疫组化法检测60例大肠癌组织及20例正常大肠黏膜组织中SS蛋白和VEGF-C蛋白的表达，采用VEGF-C受体FLT-4阳性脉管标记淋巴管计数，分析SS与大肠癌淋巴管生成、转移的关系。结果SS蛋白表达阳性率在大肠癌组及正常大肠黏膜组分别为37．7％和65％，VEGF-C在癌及正常组阳性表达率分别为60％和30％，差别均有显著性；SS表达与肿瘤分化程度、淋巴结转移、淋巴管侵犯及远处转移密切相关，与浆面膜受累无关；VEGF-C表达与肿瘤淋巴结转移，淋巴管侵犯及远处转移密切相关，而与肿瘤分化和浆面膜受累无关；大肠癌组织中SS和VEGF-C蛋白表达呈显著负相关。结论SS可能通过对VEGF-C／FLT-4信号通路的阻滞而抑制大肠癌淋巴管生成及转移。     

黏附分子CD44在大肠癌中表达的临床意义

采用免疫组化法检测121例大肠癌患者肿瘤组织的CD44v6蛋白表达。结果在大肠癌中CD44v6表达阳性率为50．4％（61／121），其中低分化者阳性率高于高分化者，淋巴结转移者高于无淋巴结转移者，Dukes分期D期者高于A期者（P〈0．05，〈0．01）。提示CD44v6表达阳性的大肠癌具有更强的浸润及淋巴结转移能力，CD44v6可作为判断大肠癌预后的一个重要指标。     

VEGFR-3在喉癌组织中的表达及临床意义

目的探讨血管内皮生长因子受体-3（VEGFR-3）在喉癌发生、发展中的作用及对预后的影响。方法选择手术切除的喉癌组织标本45份（喉癌组）,喉良性病变组织标本20份（良性组）,采用免疫组化法检测两组VEGFR-3蛋白的表达,实时荧光定量PCR法检测VEGFR-3mRNA表达;分析VEGFR-3与喉癌临床病理特征的关系,应用Log-rank检验分析VEGFR-3表达与喉癌预后的关系。结果喉癌组及良性组VEGFR-3的阳性率分别为44．4％、15．0％,VEGFR-3mRNA的相对表达量分别为10．4±2．7、2．3±0．6,两组比较P均〈0．01;VEGFR-3表达与喉癌淋巴结转移、淋巴管浸润密切相关,而与年龄、性别、肿瘤部位、组织学分级、TNM分期无关;喉癌组VEGFR-3阳性及阴性表达者平均生存时间分别为50、60个月,P〉0．05。结论VEGFR-3与喉癌淋巴结转移相关,但与预后无明显相关性。     

血小板源性生长因子受体-β在大肠癌间质中的表达及与淋巴管生成的关系

目的检测血小板源性生长因子受体－β（PDGFR—β）在大肠癌间质组织中的表达,探讨其与淋巴管生成的关系。方法应用免疫组化SP法检测90份大肠癌间质及20份正常大肠组织中PDGFR—β的表达,用D2-40标记微淋巴管,计数微淋巴管密度（LMVD）,分析PDGFR—β的表达与LMVD和临床病理参数之间的关系。结果大肠癌间质组织中PDGFR—β的表达及LMVD明显高于癌旁正常组织（P均〈0．05）;有淋巴结转移者LMVD均明显高于无淋巴转移者,PDGFR-β阳性表达者的LMVD明显高于阴性表达者。间质组织中PDGFR-β的阳性表达与淋巴结转移相关（P〈0．05）。结论PDGFR—β在大肠癌间质组织中表达与肿瘤淋巴管的形成密切相关并可促进淋巴结转移。     

泛素、Skp2、p27在大肠癌组织中的表达及意义

采用组织芯片技术制作80例大肠癌癌组织芯片。S-P免疫组织化学方法检测芯片中泛素、S期激酶相关蛋白2（Skp2）和p27表达。结果泛素表达阳性率为45．0％，Skp2阳性率为47．5％，p27阳性率为38．8％，不同组织学类型大肠癌中泛素、skp2、p27的表达未见明显差异（P〉0．05）；有淋巴结转移者Skp2阳性率显著高于无淋巴结转移者（P〈0．01），p27阳性率显著低于无淋巴结转移者（P〈0．01）。提示Skp2高表达和p27低表达与大肠癌的进展相关，Skp2和p27的表达可以作为评估大肠癌预后的参考指标；泛素和Skp2在p27的表达调控中起重要作用；应用组织芯片大规模高效检测临床组织样本是可行的，具有快速、方便、经济、准确的特点。     

VEGF-C和Podoplanin在食管癌中的表达及意义

目的 探讨食管鳞癌中VEGF-C和Podoplanin的表达及其与食管癌淋巴管生成和淋巴结转移的关系.方法 应用免疫组织化学SP法检测74例食管鳞癌标本及8例癌旁正常黏膜组织中VEGF-C和Podoplanin的表达,并对两指标表达与临床病理参数之间进行相关分析.结果 ①VEGF-C阳性率和Podoplanin阳性淋巴管密度(LVD)显著高于正常组织(P＜0.05);②VEGF-C阳性率和LVD值在淋巴结转移组明显高于无淋巴结转移组(P＜0.05);③VEGF-C阳性表达的食管癌组织中LVD与阴性表达的LVD比较差异显著(P=0.001).结论 VEGF-C和LVD是食管癌淋巴结转移和淋巴管生成的重要指标,且VEGF-C和LVD明显相关.     

Dermatomyositis with peripheral nervous system involvement: activation of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in vasculitic lesions

We describe two cases of dermatomyositis (DM) with nervous system involvement. Polyneuropathy was observed in both patients, and cerebral vasculitis was suspected in one patient. Histological examination of biopsied specimens of skeletal muscles, skin and sural nerves revealed vasculitis. Furthermore, vascular endothelial growth factor (VEGF) and VEGF receptors (VEGFRs) were overexpressed in vasculitic lesions. Although, VEGF and VEGFRs were not detected in biopsied specimens of skeletal muscle from normal subjects, they were observed in one of two patients with DM who did not exhibit neuropathy. These findings suggest the possibility that VEGF overproduction is associated with development of vasculitis in some cases of DM complicated with peripheral neuropathy.     

KDR (VEGF receptor 2) is the major mediator for the hypotensive effect of VEGF

Vascular endothelial growth factor (VEGF) exerts vasodilation-induced hypotension as a major side effect for treatment of ischemic diseases. VEGF has 2 receptor tyrosine kinases, KDR and Flt-1. Little is known about which receptor mediates VEGF-induced hypotension. To elucidate the role of each receptor in mediating hypotension, KDR-selective and Flt-1-selective mutants were used for in vitro and in vivo studies. The KDR-selective mutant induced vascular endothelial cell proliferation comparable to VEGF, whereas the Flt-1- selective mutant had no effect on proliferation. Intravenous injection of KDR-selective mutant, Flt-selective mutant, or VEGF caused a dose-related decrease in mean arterial pressure in conscious rats. The hypotensive response to KDR-selective mutant was significantly less than that to VEGF (P<0.01) but was greater than that to Flt-selective mutant (P<0.01). Similarly, VEGF and KDR-selective mutant induced more potent vasorelaxation than Flt-selective mutant or placenta growth factor that binds Flt-1 only (P<0.01), and the vasorelaxation to KDR-selective mutant was not significantly different at low concentrations but less than that to VEGF at high concentrations. The results indicate that the vasodilation and hypotensive effect of VEGF may involve both receptors, but KDR is the predominant receptor mediating this effect. Because KDR-selective mutant induced proliferation and angiogenesis similar to VEGF but was associated with 36% attenuation in hypotension, the data suggest that the KDR-selective mutant may represent an alternative treatment for ischemic diseases.     

血管内皮生长因子基因C(-634)G多态性及血浆血管内皮生长因子水平与糖尿病视网膜病变的相关性研究

目的 探讨血管内皮生长因子(VEGF)基因C(-634)G多态性及血浆中VEGF水平与糖尿病视网膜病变(DR)的关系. 方法 在昆明地区汉族人中检测96名正常对照者(NC组)和285例T2DM患者的VEGF基因C(-634)G多态性及血浆中VEGF水平. 结果 DR组CC基因型和C等位基因频率显著高于非DR(NDR)组(P均<0. 05),DR组中CC基因型者血浆VEGF水平高于CG及GG基因型者和NC组(P<0. 05或P<0. 01). 结论 VEGF基因C(-634)G多态性是昆明地区汉族T2DM患者发生DR的危险因素.     

Association of vascular endothelial growth factor (VEGF) gene polymorphism and increased serum VEGF concentration with pancreatic adenocarcinoma

Background &aimPancreatic cancer is related to high mortality rate. The vascular endothelial growth factor (VEGF) has a strong influence in tumor-related angiogenesis having association with the grade of angiogenesis and the prognosis of different solid tumors including pancreatic cancer. The present study was aimed to analyze the genotype and haplotype distribution of VEGF gene single nucleotide polymorphisms (SNPs), ?460T/C, +405G/C, +936C/T, in patients with pancreatic adenocarcinoma from South India, and the effect of these SNPs on serum VEGF level.MethodsTotal 80 patients with pancreatic adenocarcinoma and 87 controls were recruited. The genotype of VEGF gene polymorphisms was determined in both patients and controls using polymerase chain reaction-restriction fragment length polymorphism method. The serum VEGF protein was estimated by standard enzyme-linked immunosorbent assay.ResultsThe genotype, +405G/G of VEGF gene showed a significant association with the patients with pancreatic adenocarcinoma (P = 0.012, Odds ratio: 2.133), whereas no significant difference was found in the genotype distribution of SNPs, ?460C/T and +936C/T between patient and control groups (P > 0.05). Serum VEGF level was found to be significantly high in patients (1315.10 pg/Ml, SD ± 230.79) when compared to controls (591.35 pg/mL, SD ± 92.48) (P < 0.0001), which showed a strong genotype–phenotype correlation between genotype +405G/G and serum VEGF level. Further, the haplotype C-G-T showed a strong association with the disease, and no specific haplotype was associated with increased serum VEGF level.ConclusionThe polymorphism, +405G/C but not ?460T/C and +936C/T, of VEGF gene is strongly associated with pancreatic adenocarcinoma, and this SNP has significant influence on serum VEGF level.     

Binding and neutralization of vascular endothelial growth factor (VEGF) and related ligands by VEGF Trap, ranibizumab and bevacizumab

Pharmacological inhibition of VEGF-A has proven to be effective in inhibiting angiogenesis and vascular leak associated with cancers and various eye diseases. However, little information is currently available on the binding kinetics and relative biological activity of various VEGF inhibitors. Therefore, we have evaluated the binding kinetics of two anti-VEGF antibodies, ranibizumab and bevacizumab, and VEGF Trap (also known as aflibercept), a novel type of soluble decoy receptor, with substantially higher affinity than conventional soluble VEGF receptors. VEGF Trap bound to all isoforms of human VEGF-A tested with subpicomolar affinity. Ranibizumab and bevacizumab also bound human VEGF-A, but with markedly lower affinity. The association rate for VEGF Trap binding to VEGF-A was orders of magnitude faster than that measured for bevacizumab and ranibizumab. Similarly, in cell-based bioassays, VEGF Trap inhibited the activation of VEGFR1 and VEGFR2, as well as VEGF-A induced calcium mobilization and migration in human endothelial cells more potently than ranibizumab or bevacizumab. Only VEGF Trap bound human PlGF and VEGF-B, and inhibited VEGFR1 activation and HUVEC migration induced by PlGF. These data differentiate VEGF Trap from ranibizumab and bevacizumab in terms of its markedly higher affinity for VEGF-A, as well as its ability to bind VEGF-B and PlGF.     

VEGF反义寡核苷酸抑制人膀胱癌细胞VEGF表达的作用和效果

目的　观察 VEGF反义寡核苷酸对膀胱癌细胞 VEGF表达的抑制情况。方法　采用免疫组化法观察反义寡核苷酸对膀胱癌细胞VEGF表达的影响 ,并观察条件培养液对内皮细胞生长的抑制作用。结果　 VEGF反义寡核苷酸对膀胱癌细胞的 VEGF表达有明显的抑制作用。结论　 VEGF反义寡核苷酸能够抑制膀胱癌细胞的 VEGF表达 ,故对临床膀胱癌抗血管生成的治疗极具潜力和应用前景。     

Vascular endothelial growth factor (VEGF) is an autocrine growth factor for VEGF receptor-positive human tumors

Angiogenesis is required for the progression of tumors from a benign to a malignant phenotype and for metastasis. Malignant tumor cells secrete factors such as vascular endothelial growth factor (VEGF), which bind to their cognate receptors on endothelial cells to induce angiogenesis. Here it is shown that several tumor types express VEGF receptors (VEGFRs) and that inhibition of VEGF (VEGF antisense oligonucleotide AS-3) or VEGFRs (neutralizing antibodies) inhibited the proliferation of these cell lines in vitro. Furthermore, this effect was abrogated by exogenous VEGF. Thus, VEGF is an autocrine growth factor for tumor cell lines that express VEGFRs. A modified form of VEGF AS-3 (AS-3m), in which flanking 4 nucleotides were substituted with 2-O-methylnucleosides (mixed backbone oligonucleotides), retained specificity and was active when given orally or systemically in vitro and in murine tumor models. In VEGFR-2-expressing tumors, VEGF inhibition may have dual functions: direct inhibition of tumor cell growth and inhibition of angiogenesis.     

Vascular endothelial growth factor D (VEGF-D) is a ligand for the tyrosine kinases VEGF receptor 2 (Flk1) and VEGF receptor 3 (Flt4)

We have identified a member of the VEGF family by computer-based homology searching and have designated it 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. In adult human tissues, VEGF-D mRNA is most abundant in heart, lung, skeletal muscle, colon, and small intestine. Analyses of VEGF-D receptor specificity revealed that VEGF-D is a ligand for both VEGF receptors (VEGFRs) VEGFR-2 (Flk1) and VEGFR-3 (Flt4) and can activate these receptors. However, VEGF-D does not bind to VEGFR-1. Expression of a truncated derivative of VEGF-D demonstrated that the receptor-binding capacities reside in the portion of the molecule that is most closely related in primary structure to other VEGF family members and that corresponds to the mature form of VEGF-C. In addition, VEGF-D is a mitogen for endothelial cells. The structural and functional similarities between VEGF-D and VEGF-C define a subfamily of the VEGFs.     

Combination of Vascular Endothelial Growth Factor (VEGF) and Thymidine Phosphorylase (TP) to Improve Angiogenic Gene Therapy

To improve current angiogenic gene therapy with a vascular endothelial growth factor (VEGF)-encoding plasmid (Baumgartner et al. Circulation 1998; 97: 1114-23 [1]; Kusumanto et al. Fifth Annual Meeting of the American Society of Gene Therapy, Boston, 2002, Abstr. 621 [2]), we have generated a combination plasmid, encoding the VEGF gene and the thymidine phosphorylase (TP, also known as platelet-derived endothelial growth factor (PD-ECGF) or gliostatin (GLS)) gene: phVEGF165-TP.MB. Upon transfection in COS-7 cells both gene products were expressed and functional as shown by Western blots, ELISAs and bioassays. Culture supernatants of COS-7 cells transfected with this plasmid were able to induce endothelial proliferation. In an in vitro angiogenesis assay with recombinant proteins, TP was able to increase VEGF-induced tube formation. The phVEGF165-TP.MB plasmid is therefore a promising candidate for in vivo angiogenesis studies.