血管内皮细胞生长因子及其受体在结直肠癌中的表达

目的:探讨血管内皮细胞生长因子(WIGF)及其受体(FLT、FLK-1)在结直肠癌中的表达.方法:应用LSAB免疫组化技术,检测50例人结直肠癌组织VEGF、FLT、FLK-1和8因子相关抗原(FⅧRAg)的表达情况.结果:VEGF、FLT和FLK-1主要表达于侵袭性癌的浸润边缘和坏死组织周围.FLT表达于癌细胞膜和/或细胞浆,而FLK-1则只表达于细胞浆.新生血管多位于肿瘤组织的侵袭性边缘和间质浸润灶旁.结论:WIGF、FLT和FLK-1表达呈现出明显的异质性,并与肿瘤新生血管形成密切相关.综合分析VEGF及其受体的表达有助于对判断结直肠癌侵袭能力和新生血管形成的判断.     

The vascular endothelial growth factor family and its receptors

This article focuses on describing the biology of vascular endothelial growth factor (VEGF) and its receptors as well as the regulation of their expression. A thorough understanding of the VEGF system is paramount in optimizing antiangiogenic therapies as a component of antineoplastic regimens.     

The expression of vascular endothelial growth factor C and its receptors in non-small cell lung cancer.

Expression of vascular endothelial growth factor (VEGF)-C and that of its receptors were assessed in non-small cell lung cancer. Immunohistochemistry revealed positive VEGF-C expression in 38.7% (24/62) of the patients studied. A significant positive correlation was found between VEGF-C in cancer cells and VEGF receptor-3 (VEGFR-3) in vascular endothelial cells, but not between VEGF-C in cancer cells and VEGFR-2 in endothelial cells. In this cohort of lung cancer patients, VEGF-C expression was significantly associated with lymph node metastasis, lymphatic vessel invasion, and worse outcomes after the operation. Although the independent prognostic impact of VEGF-C and VEGFR-3 was not clear, VEGFR-2 expression in endothelial cells retained the independency as the prognostic indicator. In light of these findings, we conclude that VEGF-C plays an important role in lymphatic invasion/metastasis and tumour progression in non-small cell lung cancer.     

Melatonin inhibits the expression of vascular endothelial growth factor in pancreatic cancer cells

Objective:To investigate the effects of melatonin on cellular proliferation and endogenous vascular endothelial growth factor(VEGF) expression in pancreatic carcinoma cells(PANC-1).Methods:PANC-1 cells were cultured for this study.The secreted VEGF concentration in the culture medium was determined using ELISA method,VEGF production in the tumor cells was detected by immunocytochemistry,and VEGF mRNA expression was determined by RT-PCR.Results:Higher melatonin concentrations significantly inhibited cellular proliferation,with 1 mmol/L concentration exhibiting the highest inhibitory effect(P<0.01).VEGF concentrations in the cell culture supernatants and intra-cellules were all significantly reduced after melatonin(1 mmol/L) incubation(P<0.05).VEGF mRNA expression decreased markedly in a time-dependent manner during the observation period(P<0.05).Conclusions:High melatonin concentrations markedly inhibited the proliferation of pancreatic carcinoma cells.The endogenous VEGF expression was also suppressed by melatonin incubation.     

Functional significance of vascular endothelial growth factor receptors on gastrointestinal cancer cells

Vascular endothelial growth factor (VEGF) has been shown to be the major mediator of physiologic and pathologic angiogenesis. VEGF was initially thought to be an endothelial cell specific ligand, but recently, VEGF has been shown to mediate tumor cell function via activation of receptors on tumor cells themselves. Here, we review the expression patterns and binding profiles of the VEGF receptors and their ligands on gastrointestinal tumor cells. Furthermore, we describe the current knowledge in regards to the function of these receptors on tumor cells. Elucidating the function of VEGF receptors on tumor cells should help us to better understand the potential mechanisms of action of anti-VEGF therapies.     

Pancreatic carcinoma cells express neuropilins and vascular endothelial growth factor, but not vascular endothelial growth factor receptors

BACKGROUND: Neuropilins (NRPs) are characterized as coreceptors of vascular endothelial growth factor (VEGF). In the current study, the authors assessed the expression of NRPs, VEGF, and vascular endothelial growth factor receptors (VEGFRs), as well as VEGF-induced cell proliferation, in pancreatic carcinoma cell lines and tissue specimens. METHODS: Human pancreatic carcinoma cell lines (Panc-1 and MIA PaCa-2), normal human pancreatic ductal epithelial cells (HPDE), and human umbilical vein endothelial cells (HUVECs) were cultured. Human pancreatic adenocarcinoma tissue specimens were also studied. Expression levels of NRPs, VEGFRs, and VEGF were determined by real-time polymerase chain reaction analysis and immunostaining. Cell proliferation was examined using a [3H]thymidine incorporation assay. RESULTS: Both NRP-1 and NRP-2 were expressed in Panc-1 cells, HPDE cells, and HUVECs but were expressed minimally in MIA PaCa-2 cells. Panc-1 expressed 30 times more NRP-1 mRNA than NRP-2 mRNA. NRP-1 levels in Panc-1 cells were 5.3 times higher than in HPDE cells but were similar to NRP-1 levels in HUVECs. NRP-2 levels in Panc-1 cells were similar to NRP-2 levels in HPDE cells but lower than NRP-2 levels in HUVECs. Expression of all three VEGFRs was observed only in HUVECs. However, VEGF mRNA was detected in all cell types except for HUVECs. NRP-1 immunoreactivity levels were much higher than NRP-2 immunoreactivity levels in Panc-1 and human pancreatic adenocarcinoma tissue specimens, whereas VEGFRs were not detected in either of these two settings. In response to VEGF165, [3H]thymidine incorporation in Panc-1 cells increased significantly (by 61%; P < 0.01). A monoclonal antibody against human NRP-1 significantly blocked VEGF-induced cell proliferation in Panc-1 cells. CONCLUSIONS: The pancreatic carcinoma cell line Panc-1 and adenocarcinoma tissue specimens expressed high levels of NRP-1 and VEGF, but not VEGFRs, and exogenous VEGF significantly increased NRP-1-mediated, but not VEGFR-mediated, Panc-1 cell proliferation. These data suggested that NRP-1 may be involved in the pathogenesis of pancreatic carcinoma.     

肿瘤细胞系血管内皮生长因子及其受体共表达的研究

背景与目的：血管内皮生长因子（vascular endothelial growth factor,VEGF)旁分泌在肿瘤血管新生中的作用已得到证实,但其自分泌作用尚不清楚。本研究的目的是分析VEGF及其受体（Flt-1和KDR）基因在恶性肿瘤细胞系中的共表达。方法：以看家基因为内标,采用半定量逆转录－聚合酶链反应分析VEGF及其受体基因在30种肿瘤细胞系和4种内皮细胞中的表达水平。结果：在29种肿瘤细胞系和3种内皮细胞系检测到中度以上的VEGF基因表达,而人脐静脉内皮细胞仅有低表达;Flt-1基因表达分别见于50％（6／12）的血液肿瘤,28％（5／18）的实体瘤细胞和2种内皮细胞;仅在16．7％（2／12）的血液肿瘤,33．3％（6／18）实体瘤细胞和2种内皮细胞检测到KDR基因表达;而ECV304细胞并无Flt-1或KDR基因的表达。结论：VEGF基因高表达是肿瘤细胞的重要特征,而VEGF及其受体共表达表明肿瘤细胞系中存在自分泌途径。     

宫颈癌中VEGF-C及其受体mRNA的表达及其临床意义

背景与目的：血管内皮生长因子C（vascular endothelial growth factor C，VEGF-C）是一个较为特异的淋巴内皮细胞生长刺激因子，通过其受体VEGFR-2（KDR）及受体VEGFR-3（Flt-4）分别作用于血管和淋巴管上皮促进肿瘤的生长和转移。研究宫颈癌组织及癌旁组织中血管内皮生长因子C及其受体mRNA的表达情况，分析其在肿瘤转移中的作用。方法：采用RT-PCR法分析48例新鲜宫颈癌组织及癌旁组织标本中VEGF-C／KDR／Flt-4mRNA的表达情况，并分析其与各临床病理参数之间的关系。结果：48例宫颈癌组织与瘤旁组织中的VEGF-C及其受体mRNA表达率明显高于正常宫颈组织，肿瘤组织中VEGF-C mRNA表达与flt-4 mRNA的表达呈显著正相关，癌旁组织（PT）中VEGF-C mRNA表达与KDR mRNA的表达显著相关（P〈0．001）。宫颈癌组织的VEGF-C、KDR或Flt-4 mRNA表达与肿瘤的病理类型及临床病理分期无明屁的相关；而与肿瘤的病理分化程度、淋巴结转移、肿瘤直径、深肌层浸润间差异有显著性（P〈0．05）。结论：VEGF-C可能在宫颈癌转移尤其是淋巴转移中起重要作用，是一个反映患者淋巴转移和预后的良好指标。     

VEGF-D及其受体Flt-4在胰腺癌中的表达及意义

目的：探讨血管内皮生长因子D（vascular endothelial growth factor D，VEGF—D）及其受体Flt-4（fms—like tyrosine kinase-4）在胰腺癌中的表达及其与胰腺癌临床病理特征及预后的关系。方法：利用免疫组织化学染色法检测48例胰腺癌组织、32例癌旁胰腺组织及13例正常胰腺组织中VEGF—D及Flt-4蛋白的表达情况，结合临床病理特征及预后对其进行统计学分析。结果：VEGF-D及Flt-4在胰腺癌组织中的表达率显著低于癌旁胰腺组织中的表达率（P〈0．05）并显著高于正常胰腺组织中的表达率（P〈0．05），胰腺癌组织中有淋巴结转移的VEGF-D及Flt-4的表达率显著高于无淋巴结转移组的表达率（P〈0．05），VEGF-D阳性组和Flt-4阳性组患者的中位生存期及1、2、3年生存率均显著低于阴性组（P〈0．05）。结论：VEGF-D及Flt-4可促进胰腺癌淋巴结转移的发生，并可作为判断胰腺癌患者预后的预测因素。     

VEGF及其受体在多发性骨髓瘤中的表达及意义

背景与目的:骨髓新生血管形成在多发性骨髓瘤(multiple myeloma,MM)的发生、发展和预后中起着重要的作用,血管内皮生长因子(vascular endothelialgrowth factor,VEGF)在此过程中扮演了关键角色.本研究旨在研究VEGF及其受体在MM中的表达,分析其与MM发生、发展的关系.方法:采用RT-PCR的方法检测35例MM患者、16例非肿瘤患者(下称对照组)以及KM3细胞株中VEGF及其受体Flt-1和KDR的表达,并分析阳性率以及表达相对含量在MM患者和非肿瘤患者、MM不同分期间的差异.结果:VEGF和Flt-1基因在MM组的阳性率(62.9%和80.0%)显著高于对照组(18.8%和31.3%)(P＜0.01),VEGF在两组中的表达水平为0.41±0.19 vs.0.06±0.01(P＜0.05),Flt-1为0.60±0.33 vs.0.08±0.03(P＜0.01);VEGF基因在初治组和复发/难治MM组的阳性率为66.7%vs.60.9%(P＞0.05),Flt-1基因为83.3%vs.78.3%(P＞0.05);VEGF在Ⅱ期和Ⅲ期MM的阳性率为50%vs.73.7%(P＞0.05),Flt-1为81.3%vs.78.9%(P＞0.05);复发/难治MM组的VEGF和Flt-1基因的表达水平明显高于初治组(0.49±0.20 vs.0.28±0.04,P＜0.05;0.70±0.38 vs.0.41±0.06,P＜0.05),Ⅲ期MM患者VEGF和Flt-1基因的表达水平明显高于Ⅱ期患者(0.48±0.19 vs.0.28±0.09,P＜0.05;0.75±0.35 vs.0.41±0.21,P＜0.05).KDR仅在3例MM患者中检出,对照组未检出.结论:VEGF和Flt-1在MM中高表达,并与疾病进展相关.     

Vascular endothelial growth factor and two types of its receptors in breast cancer

Levels of VEGF, VEGFR-1 and VEGFR-2 were determined by enzyme immunoassay in tumor and adjacent normal tissue samples from 39 breast cancer patients. Those parameters were significantly higher in tumor tissue. Direct correlations were established between VEGF and VEGFR-1, on the one hand, and VEGF and VEGFR-2, on the other. VEGF expression in breast tumor was relatively higher at earlier stages of tumor growth. VEGF and VEGFR-1 expression was consistently higher in progesterone receptor negative tumors.     

Inhibited growth of colon cancer carcinomatosis by antibodies to vascular endothelial and epidermal growth factor receptors

Vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) regulate colon cancer growth and metastasis. Previous studies utilizing antibodies against the VEGF receptor (DC101) or EGF receptor (C225) have demonstrated independently that these agents can inhibit tumour growth and induce apoptosis in colon cancer in in vivo and in vitro systems. We hypothesized that simultaneous blockade of the VEGF and EGF receptors would enhance the therapy of colon cancer in a mouse model of peritoneal carcinomatosis. Nude mice were given intraperitoneal injection of KM12L4 human colon cancer cells to generate peritoneal metastases. Mice were then randomized into one of four treatment groups: control, anti-VEGFR (DC101), anti-EGFR (C225), or DC101 and C225. Relative to the control group, treatment with DC101 or with DC101+C225 decreased tumour vascularity, growth, proliferation, formation of ascites and increased apoptosis of both tumour cells and endothelial cells. Although C225 therapy did not change any of the above parameters, C225 combined with DC101 led to a significant decrease in tumour vascularity and increases in tumour cell and endothelial cell apoptosis (vs the DC101 group). These findings suggest that DC101 inhibits angiogenesis, endothelial cell survival, and VEGF-mediated ascites formation in a murine model of colon cancer carcinomatosis. The addition of C225 to DC101 appears to lead to a further decrease in angiogenesis and ascites formation. Combination anti-VEGF and anti-EGFR therapy may represent a novel therapeutic strategy for the management of colon peritoneal carcinomatosis.     

Circulating vascular endothelial growth factor and interferon-gamma-inducible protein-10 levels in pancreatic cancer during chemotherapy

BACKGROUND: Chemotherapeutic anticancer properties are thought to derive from apoptosis pathway activation and/or cell division arrest, but animal models have also evidenced anti-angiogenic activity in some agents. PATIENTS AND METHODS: The impact of gemcitabine, irinotecan and oxaliplatin + 5-FU upon the serum markers vascular endothelial growth factor (VEGF) (pro-angiogenic) and IFN-gamma-inducible protein (IP)-10 (anti-angiogenic) was evaluated by ELISA in locally advanced and/or metastatic cancer versus clinical efficacy and survival. RESULTS: Patients had higher serum levels of both markers versus controls. No objective response to therapy was observed and no significant difference in either marker occurred during the first month of chemotherapy; analysis by survival showed slight transient VEGF decrease in longer survivors on day 14 and slight increase on day 28 in shorter survivors, who had baseline median IP-10 levels above longer survivors, diverging on day 14 (decrease and increase, respectively). Both groups were below baseline at day 28. Changes in IP-10 were not significant. CONCLUSION: These preliminary results provide a rationale for exploring whether continuous or frequent administration of some anti-neoplastic agents may elicit a global anti-angiogenic activity, and whether different administration schedules of the same drug could have a synergistic or an antagonistic effect, which obviously would need to be taken into account in determining combinations with new agents targeting angiogenesis.     

食管癌患者血清VEGF水平及其临床意义

目的检测食管癌患者血清中血管内皮生长因子（VEGF）水平并探讨其临床意义。方法用ELISA方法检测38例食管癌和82例正常人群血清中VEGF165的浓度。结果正常健康人血清中存在VEGF165,其正常值为0．188μg／L,食管鳞癌者血清中VEGF水平明显高于正常对照组,血清VEGF水平随着食管临床病理分期的进展而逐步升高。结论VEGF血清水平在一定程度上反映食管癌患者的临床病理分期,有利于对治疗措施的选择和预后的判断。     

Concomitant over-expression of activin/inhibin β subunits and their receptors in human pancreatic cancer

Activins and inhibins belong to the transforming growth factor-β (TGF-β) superfamily of multifunctional cytokines that bind to transmembrane receptors with serine/threonine kinase activity. In this study, we characterized the levels of expression of 3 activin/inhibin subunits (βA, βB, α), and 2 type I and type II activin receptors (actRI/Ib, actRII/IIb) in pancreatic cancer cell lines and in human pancreatic tissues. In addition, we assessed the growth responsiveness to activin A in these cell lines. All 6 cell lines (ASPC-1, CAPAN-1, COLO-357, MIA-PaCa-2, PANC-1 and T3M4) expressed the activin/inhibin βA subunit, whereas expression levels of the activin/inhibin βB and α subunits were undetectable. Furthermore, actRI, actRII and actRIIb were expressed in all cell lines and actRIb mRNA was evident in ASPC-1, CAPAN-1, COLO-357 and PANC-1 cells. CAPAN-1 and COLO-357 cells were growth-stimulated by activin A in the presence of 10% serum, whereas the other cell lines were resistant to activin A. In contrast, in serum-free medium activin A inhibited the growth of CAPAN-1, COLO-357 and MIA-PaCa-2 cells. Pancreatic cancer samples markedly over-expressed the activin/inhibin βA subunit, whereas the βB subunit was only moderately increased in comparison to normal pancreatic samples. Pancreatic cancer tissues also markedly over-expressed actRI, actRIb and actRII. By in situ hybridization, activin/inhibin βA, actRI, actRIb and actRII were strongly expressed in diffuse infiltrative and duct-like cancer cells. Both the ligand and its receptors were often co-expressed in these cells. Together, our findings suggest that activin A may participate in autocrine activation of pancreatic cancer cells in vivo. Int. J. Cancer 77:860–868, 1998.© 1998 Wiley-Liss, Inc.     

青蒿琥酯对急性单核细胞白血病SHI-1细胞株血管内皮生长因子及其受体表达的影响

 目的　研究青蒿琥酯对急性单核细胞白血病SHI-1细胞株血管内皮生长因子（VEGF）及其受体（VEGFR）的影响。方法　酶联免疫吸附法检测非细胞毒性浓度（5、10、20 ng／ml）青蒿琥酯作用SHI-1细胞后培养上清液VEGF浓度,流式细胞术检测有或无青蒿琥酯作用时,SHI-1细胞表面VEGFR-1及VEGFR-2阳性表达率。结果　培养24、48 h后,无青蒿琥酯作用的 SHI-1细胞培养上清液VEGF质量浓度分别为（980.3±2.2）、（982.4±2.3）pg／ml,VEGFR-1表达率分别为（5.40±3.11）%和（4.45±2.85）%,VEGFR-2表达率分别为（13.90±2.26）%和（13.95±1.96）%。 5、10 、20 ng／ml青蒿琥酯作用24 h后,SHI-1细胞培养上清液VEGF质量浓度分别为（234.6±1.8）、（114.9±1.6）、（108.8±1.5）pg／ml,作用48 h后分别为（62.3±1.7）、（60.9±1.6）、（32.7±1.7）pg／ml,与培养相同时间无青蒿琥酯组相比,VEGF浓度明显下降（均P＜0.05）,且相同浓度青蒿琥酯作用24 h与48 h间差异亦有统计学意义（均P＜0.05）。5、10 、20 ng／ml青蒿琥酯作用24 h,VEGFR-1阳性率分别为（4.30±2.21）%、（4.20±1.37）%和（3.90±1.86）%,作用48 h后分别为（3.80±2.87）%、（3.60±1.73）%和（3.00±1.82）%,相同作用时间不同浓度青蒿琥酯组间及相同浓度作用不同时间组间VEGFR-1阳性率差异均无统计学意义（均P＞0.05）;作用24 h后,SHI-1细胞VEGFR-2阳性率分别为（4.40±1.15）%、（3.10±0.68）%和（1.10±0.72）%,作用48 h后分别为（3.00±1.68）%、（2.20±0.93）%和（0.60±0.92）%,3个不同浓度青蒿琥酯作用相同时间后VEGFR-2表达率降低（均P＜0.05）,相同浓度作用24与48 h间差异均无统计学意义（均P＞0.05）。结论　SHI-1细胞株高分泌VEGF,青蒿琥酯可下调VEGF分泌及VEGFR-2的表达,而对VEGFR-1表达的调节作用不显著。     

New therapeutic directions for advanced pancreatic cancer: targeting the epidermal growth factor and vascular endothelial growth factor pathways

In advanced pancreatic cancer, single-agent gemcitabine became the standard therapy approximately 10 years ago. Subsequently, combinations of gemcitabine with fluorouracil, cisplatin, irinotecan, oxaliplatin, or pemetrexed produced no clear survival benefit. Among the newer approaches, targeting human epidermal growth factor receptor (HER-1/EGFR) shows promise. The U.S. Food and Drug Administration recently approved erlotinib (a HER-1/EGFR tyrosine kinase inhibitor) combined with gemcitabine for the first-line treatment of advanced pancreatic cancer. This combination showed a statistically significant survival benefit over gemcitabine alone in locally advanced or metastatic disease (the median overall survival time was 6.24 months versus 5.91 months; hazard ratio, 0.82; p = .038); however, the clinical significance of this survival difference has been questioned. Additionally, a large phase III trial where the addition of cetuximab (an anti-HER-1/EGFR monoclonal antibody [mAb]) to gemcitabine failed to result in a longer overall survival time than with gemcitabine alone has been reported. Targeting vascular endothelial growth factor (VEGF) with bevacizumab (a recombinant, humanized IgG1 mAb that binds to VEGF) in combination with gemcitabine was investigated in a phase II trial, with promising outcomes that were unfortunately not supported by a subsequent phase III study. While the future treatment of pancreatic cancer may be influenced by the potential of certain biomarkers to predict better response to molecular-targeted therapies, allowing individualization of patient therapy, there are currently no clear candidates, and this remains an interesting area for further investigation.