Chlamydia pneumoniae stimulates the proliferation of HUVEC through the induction of VEGF by THP-1

Chlamydia pneumoniae, a gram-negative bacterium, is an important human intracellular pathogen; studies of C. pneumoniae pathogenesis have shown that the organism can infect many cell types associated with both respiratory and vascular sites, including arterial smooth muscle cells, macrophages and vascular endothelium. Recently, the recognition of atherosclerosis as an inflammatory disease in its genesis, progression and ultimate clinical manifestations has created an interesting area of vascular research. We assessed the hypothesis that growth factors from THP-1 macrophages infected with C. pneumoniae are involved in the regulation of cell proliferation in HUVEC. The induction of these factors were dependent on time of infection, as medium harvested 48 h after infection had a greater activity than media harvested at 12 or 24 h after infection. Heat-killed C. pneumoniae produced similar results to those of live bacteria. In addition, conditioned medium filtered sterile from infected macrophages induced the proliferation of HUVEC, thus demonstrating its angiogenic potential. Moreover, pretreatment of macrophages with cytochalasin D, a phagocytosis inhibitor, yielded almost comparable results, suggesting that bacterium cell-attachment is sufficient for VEGF, IL-1beta and IL-8 induction. Further studies are necessary to elucidate the biological role of chlamydial involvement in the complex and mutifactored processes of angiogenesis and possibly contribute to the development of therapeutic strategies.     

辛伐他汀对脐静脉内皮细胞金属基质蛋白酶9表达的影响

目的观察辛伐他汀对脐静脉内皮细胞（human umbilical vein endothelial cell,HUVEC）金属基质蛋白酶9（matrix metalloproteinase-9,MMP-9）表达的影响。方法采用逆转录聚合酶链反应及蛋白质免疫印迹分析检测MMP-9mRNA转录和蛋白水平表达,观察辛伐他汀不同浓度及不同孵育时间HUVECMMP-9表达的影响。结果辛伐他汀呈浓度和时间依赖性减低HU-VEC的MMP-9mRNA转录和蛋白水平的表达。结论辛伐他汀可抑制HUVE CMMP-9表达,防治动脉粥样硬化。     

Camptothecin suppresses expression of matrix metalloproteinase-9 and vascular endothelial growth factor in DU145 cells through PI3K/Akt-mediated inhibition of NF-κB activity and Nrf2-dependent induction of HO-1 expression

Though camptothecin (CPT) possesses potent anti-inflammatory, immunomodulatory, anticancerous, and antiproliferative effects, little is known about the mechanism by which CPT regulates the expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF). Therefore, the current study aimed to investigate the effects of CPT on the expression of MMP-9 and VEGF, which are important factors for the invasion of tumors. In vitro application of CPT resulted in a slight inhibition of cell proliferation and a significant reduction in the matrigel invasion of DU145 cells. Treatment with CPT also downregulated phorbol-12-myristate-13-acetate (PMA)- and tumor necrosis factor-α (TNF-α)-induced MMP-9 and VEGF expression by inhibiting nuclear factor-κB (NF-κB) activity. Downregulation of phosphoinositide 3-kinase (PI3K)/Akt phosphorylation in response to CPT was revealed as an upstream pathway regulating the expression of MMP-9 and VEGF accompanying the inhibition of NF-κB activity. We further confirmed that CPT inhibits PMA-induced MMP-9 and VEGF expression by upregulating nuclear factor-erythroid related factor-2 (Nrf2)-mediated heme oxygenase-1 (HO-1) induction. Taken together, these data indicate that CPT inhibits the invasion of cancer cells accompanied by suppression of MMP-9 and VEGF production by suppressing the PI3K/Akt-mediated NF-κB pathway and enhancing the Nrf2-dependent HO-1 pathway, suggesting that CPT may be a good candidate to inhibit MMP-9 and VEGF expression.     

Effects of DNMT and MEK inhibitors on the expression of RECK, MMP-9, -2, uPA and VEGF in response to arsenite stimulation in human uroepithelial cells

It has been shown that the ingestion of arsenic-contaminated drinking water is closely correlated with risk of several cancers. The mechanism of arsenic-induced carcinogenesis is still unclear. The RECK, MMP-9, -2, uPA and VEGF are the most common dysregulation in human tumors and cancer cell lines. However, the effect of arsenite on these markers expression and the molecular mechanism are still unclear. The purpose of the study was to investigate the relationship between the expression of RECK, MMP-9, -2, uPA and VEGF in arsenite-treated human and rat uroepithelial cells. In addition, we also observed and compared the expression of these markers in urothelial carcinoma (UC) from Blackfoot disease (BFD) areas and non-Blackfoot disease (non-BFD) areas. We analyzed the arsenite causing cell proliferation, RECK, MMP-9, -2, uPA and VEGF expression by Western blotting, immunocytochemistry (ICC), RT-PCR, and gelatin zymography. We demonstrated the effect of arsenite on methylation status of RECK promoter as determined by using methylation-specific PCR (MSP). Our results show that arsenite downregulation of RECK is caused by epigenetic inactivation via promoter hypermethylation, and that levels of MMP-9, -2, uPA and VEGF were increased in human uroepithelial cells (SV-HUC-1). However, when the cells were pretreated with inhibitors (5-aza-CdR or U0126) for 24 h, the effects of arsenite on RECK, MMP-9, -2, uPA and VEGF expression were suppressed. Indeed, we also found significant differences between the expression of RECK, MMP-9, -2, uPA and VEGF in UC from the BFD areas and non-BFD areas (p = 0.006, 0.007, 0.003, <0.001 and 0.001 respectively), as detected by immunohistochemistry (IHC). In in vivo study, our results showed the RECK protein expression was reduced and the expression of MMP-9, -2, uPA and VEGF increased in arsenite treatment groups. In conclusion, our results support the notion that arsenite might cause the histologic changes, RECK, MMP-9, -2, uPA and VEGF dysregulation through epigenetic inactivation and ERK1/2 activation in SV-HUC-1 cells. These findings may provide a better understanding of the urothelial carcinogenesis of arsenite.     

欧芹素乙对人脐静脉内皮细胞的保护作用及对血管内皮生长因子表达的影响

目的:研究欧芹素乙对内皮细胞的保护作用;溶血磷脂酰胆碱(LPC)对人脐静脉内皮细胞株HUVEC细胞中血管内皮生长因子(VEGF)表达的影响以及欧芹素乙的影响.方法:应用四唑盐(MTT)法检测溶血磷脂酰胆碱对HUVEC细胞的毒性作用及欧芹素乙的保护作用;应用基础酶联免疫吸附试验(ELISA)检测各组条件培养基中VEGF蛋白含量;采用RT-PCR法及Reahime PCR方法检测溶血磷脂酰胆碱对VEGF mRNA的表达及欧芹素乙的影响.结果:MTT检测结果显示,溶血磷脂酰胆碱对HUVEC细胞具有较强的生长抑制作用,而欧芹素乙对溶血磷脂酰胆碱所致的细胞增殖抑制具有较好的保护作用.ELISA结果显示,HUVEC细胞暴露于溶血磷脂酰胆碱后,VEGF蛋白含量明显升高;加入欧芹素乙后剂量依赖性地降低VEGF蛋白的表达.RT-PCR结果显示,溶血磷脂酰胆碱可以增加3种VEGF异构体的转录水平,其中VEGF165的表达显著增加,欧芹素乙可剂量依赖性地抑制溶血磷脂酰胆碱引起的VEGF mRNA的高表达.结论:欧芹素乙对溶血磷脂酰胆碱引起的细胞损伤有明显的保护作用;欧芹素乙可抑制溶血磷脂酰胆碱所诱导的HUVEC细胞中VEGF蛋白及VEGF mRNA的高表达,对内皮细胞起到保护作用.     

Insulinotropic compounds decrease endothelial cell survival

ObjectivesHyperglycemia induces damage of vascular endothelial cells leading to diabetic complications. We investigated the effects of insulinotropic compounds and elevated glucose on endothelial cells in the absence or presence of vascular endothelial growth factor (VEGF).ResultsHuman umbilical vein endothelial cells (HUVECs) were treated with glibenclamide, repaglinide and insulinotropic imidazolines at high glucose concentration in the presence or absence of VEGF and viability, proliferation and nitric oxide production were measured. Hyperglycemia inhibited pro-survival effects of VEGF on endothelial cells. Glibenclamide and repaglinide decreased HUVEC viability at elevated glucose concentration in the absence but not in the presence of VEGF, without affecting HUVEC proliferation. Repaglinide also had some positive influence on HUVEC function elevating NO production in the presence of VEGF. Imidazolines showed different activities on endothelial cell survival. Efaroxan diminished HUVEC viability at elevated glucose concentration in the presence, however not in the absence of VEGF, while RX871024 decreased HUVEC survival regardless of the presence of VEGF.Significance of the studyOur data demonstrate an important interplay between the actual insulinotropic compounds, VEGF and ambient glucose concentration affecting the survival of the vascular endothelial cells. Consequently, this interplay needs to be taken into consideration when designing novel oral antidiabetic compounds.     

Decursin inhibits induction of inflammatory mediators by blocking nuclear factor-kappaB activation in macrophages

In the course of screening inhibitors of matrix metalloproteinase (MMP)-9 induction in macrophages, we isolated decursin, a coumarin compound, from the roots of Angelicae gigas. As a marker for the screening and isolation, we tested expression of MMP-9 in RAW264.7 cells and THP-1 cells after treatment with bacterial lipopolysaccharide (LPS), the TLR-4 ligand. Decursin suppressed MMP-9 expression in cells stimulated by LPS in a dose-dependent manner at concentrations below 60 microM with no sign of cytotoxicity. The suppressive effect of decursin was observed not only in cells stimulated with ligands for TLR4, TLR2, TLR3, and TLR9 but also in cells stimulated with interleukin (IL)-1beta, and tumor necrosis factor (TNF)-alpha, indicating that the molecular target of decursin is common signaling molecules induced by these stimulants. In addition to the suppression of MMP-9 expression, decursin blocked nitric oxide production and cytokine (IL-8, MCP-1, IL-1beta, and TNF-alpha) secretion induced by LPS. To find out the molecular mechanism responsible for the suppressive effect of decursin, we analyzed signaling molecules involved in the TLR-mediated activation of MMP-9 and cytokines. Decursin blocked phosphorylation of IkappaB and nuclear translocation of NF-kappaB in THP-1 cells activated with LPS. Furthermore, expression of a luciferase reporter gene under the promoter containing NF-kappaB binding sites was blocked by decursin. These data indicate that decursin is a novel inhibitor of NF-kappaB activation in signaling induced by TLR ligands and cytokines.     

重组VEGF165基因腺相关病毒的包装和鉴定

目的包装重组VEGF165基因的无致病性腺相关病毒,以其作为基因载体感染HUVEC细胞并检测其表达,并在体外检测病毒生物学活性。方法从含有VEGF165基因的pET-32a（＋）-VEGF165原核表达载体中扩增出VEGF165片段,构建重组骨架质粒pAAV-VEGF165。将此质粒和对照质粒pAAV-GFP分别与腺相关病毒包装质粒pAAV-RC、辅助质粒pAAV-Helper用钙-磷共转法转染HEK293细胞,通过同源重组分别产生rAAV-VEGF165和rAAV-GFP重组腺相关病毒。通过real-timePCR法测定病毒滴度后,转染HUVEC细胞并检测其表达,并通过感染HUVEC来检测其促增殖作用。结果扩增出的VEGF165片段成功构建至重组骨架质粒中,pAAV-VEGF165经双酶切和测序鉴定正确。病毒包装效率达90%以上,收获纯化rAAV-VEGF165病毒滴度达6.0×1010pfu/mL。重组腺病毒rAAV-VEGF165能够感染HUVEC细胞并得到显著性表达;与未转染组和GFP组相比,能够显著促进HUVEC细胞的增殖。结论我们成功包装了重组腺相关病毒rAAV-VEGF165,它能够感染HUVEC细胞并高表达VEGF165蛋白,并具有促增殖作用,这为进一步开展VEGF165基因的基因靶向治疗奠定了基础。     

VEGF及其受体与MMP-9在卵巢上皮性癌中的表达

目的：研究血管内皮生长因子（VEGF）及其受体、MMP-9的表达在卵巢上皮性癌侵袭转移方面的作用。方法：免疫组化SABC法检测VEGF及其受体、MMP-9蛋白的表达，并对VEGF、MMP-9蛋白表达及其相互关系进行秩和检验、Spearman等级分析，采用Kaplan-Meier分析进行生存分析。结果：VEGF广泛表达于卵巢上皮性癌的瘤细胞胞浆中，疾病晚期、分化差、腹水量多、残余瘤组织多的患者表达增强；VEGF受体表达于卵巢癌血管内皮细胞的胞浆．尤其是靠近癌巢处的内皮细胞；MMP-9表达于卵巢上皮性癌的间质细胞，疾病晚期、分化差的患者表达增强：VEGF与MMP-9在卵巢上皮性癌中的表达呈正相关。结论：VEGF与MMP-9在血管生长和局部侵袭及远处转移中可能有协同作用。     

川崎病急性期血清对血管内皮MMP-2、MMP-9和TNF-α分泌的影响及IVIG的干预机制探讨

目的：观察川崎病（Kawasaki disease,KD）患儿急性期血清对脐静脉内皮细胞（HUVEC）分泌基质金属蛋白酶-2（matrix metalloproteases-2,MMP-2）、基质金属蛋白酶-9（matrix metalloproteases-9,MMPO）和TNF-α的影响及人血丙种球蛋白（IVIG）对上述过程的调节作用。方法：HUVEC培养分为正常血清组（C组）、发热血清组（F组）、川崎病冠脉无损害血清组（NAC组）、川崎病冠脉无损害血清4-IVIG组（NACI组）、川崎病冠脉损害血清组（AC组）、川崎病冠脉损害血清＋WIG组（ACI组）。每组分别培养12h后吸取上清液,应用ELISA法测定细胞上清液MMP-2、MMP-9和TNF-α水平。结果：经川崎病患儿血清作用后的HUVEC培养上清液中MMP．2、MMP．9和TNF-α水平升高明显,与C组、F组相比差异有统计学意义（P均〈0．01）,其中AC组更高,与NAC纽相比差异有统计学意义（P〈0．01）;F组上清液中MMP-2、MMP-9和TNF-α水平虽然低于川崎病组,但仍高于C组（P〈0．01）。经丙种球蛋白干预后,AC组、NAC组MMP-2、MMP-9和TNF-α水平下降,差异有统计学意义（P均〈0．01）。HUVEC培养上清液中MMP-2、MMP-9与TNF-α含量之间呈显著性正相关（r=0．839,0．765,P〈0．01）。结论：川崎病患儿急性期血清能诱导HUVEC分泌MMP-2、MMP-9和TNF-α增加,MMP-2、MMP-9和TNF-α可能参与川崎病患儿冠脉损害的发生发展;IVIG防治冠脉损害的机制,可能与抑制内皮细胞分泌MMP-2、MMP-9和TNF-α有关。     

A mechanistic study of colon cancer growth promoted by cigarette smoke extract

Substantial evidence indicates that significant exposure to cigarette smoke is associated with an elevated risk for colorectal cancer. However, the mechanisms underlying the causal relationship between cigarette smoking and colorectal cancer remain to be investigated. Our previous study showed that cigarette smoke promotes the formation of inflammation-associated colonic adenoma in mice through an angiogenic pathway. Therefore, in the present study, we used the human colon adenocarcinoma cell line, SW1116, and human umbilical vascular endothelial cells (HUVECs) to elucidate the possible mechanisms in vitro. Results showed that cigarette smoke extract enhanced cell proliferation and the expression of 5-lipoxygenase (5-LOX), vascular endothelium growth factor (VEGF), matrix metalloproteinases (MMPs) 2 and 9 in SW1116 cells. Inhibition of 5-LOX decreased cell proliferation and expressions of VEGF, MMP-2 and MMP-9 induced by cigarette smoke extract. In addition, cigarette smoke extract indirectly stimulated HUVEC proliferation, a biological activity closely related to angiogenesis during tumor growth. This was again blocked by the 5-LOX inhibitor. Taken together, the results of the present study demonstrate the central role of 5-LOX and its relationship with angiogenic mediators in the actions of cigarette smoke in the promotion of angiogenesis during colon cancer growth.     

“Real life” polycyclic aromatic hydrocarbon (PAH) mixtures modulate hCG,hPL and hPLGF levels and disrupt the physiological ratio of MMP-2 to MMP-9 and VEGF expression in human placenta cell lines

Using JEG-3 and BeWo cells, we examined the effect of “real life” mixtures of polycyclic aromatic hydrocarbons (PAHs), at doses reported in maternal blood (Mix I) and in placental tissue (Mix II), on human chorionic gonadotropin (hCG), placental lactogen (hPL) and placental growth factor (hPLGF) secretion, protein expression and immunolocalization. Additionally, the action of PAH mixtures on basal and hormone-stimulated matrix metalloproteinase-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) protein expression was evaluated. Under basal conditions, the PAH mixtures increased hCG and decreased hPLGF levels in both cell lines, while hPL expression was stimulated in JEG-3 and inhibited in BeWo. There was no effect on the MMP-2/MMP-9 ratio or VEGF expression. In hormone-stimulated cells, PAH mixtures changed the MMP-2/MMP-9 ratio in JEG-3 cells in favor of MMP-9, while in BeWo MMP-2 was favored. The effect on VEGF expression was cell specific and dependent on the mixture. In hCG-treated cells, only Mix II inhibited VEGF expression in JEG-3 cells. Neither PAH mixtures affected this protein in BeWo cells. In hPL-treated cells, Mix I had a stimulatory effect in JEG-3 cells, while Mix II exerted an inhibitory effect in BeWo cells. In hPLGF-treated cells, Mix II decreased in JEG-3 cells, but in BeWo cells, both mixtures increased VEGF expression. Considering that the evaluated protein hormones play crucial roles in angiogenesis and neovascularization in the placenta, “real life” PAH mixtures by disrupting protein hormones levels, the MMP-2/MMP-9 ratio and VEGF expression can lead to insufficiency and many pregnancy-related disorders.     

The effect of δ-opioid agonists on intracellular calcium level in MOLT-4 T-cell line

δ-opioid agonists were reported to affect T cell functions: proliferation, cytotoxicity, cytokine production. Changes in intracellular calcium level are important in T-cell activation. In this study the effect of the synthetic δ-opioid agonist DADLE and an endogenous δ-opioid pentapeptide Met-enkephalin on the intracellular calcium level in human T-lymphoblastic leukemia MOLT-4 cells is reported. Intracellular calcium level was monitored using QUIN 2-AM as a fluorescent dye in MOLT-4 cells after short treatment (2 and 15 min) with DADLE and Met-enkephalin. DADLE (10^-8M) mildly (average 28%) decreased the intracellular calcium level after 1 min treatment. The suppressive effect of DADLE (10^-8M) on the intracellular calcium level was enhanced by longer (15 min) treatment of MOLT-4 cells (average 40%). Met-enkephalin (10^-9M - 10^-7M) decreased (average 33 %) the intracellular calcium level after 2 min treatment (average 33% - 37%). However, Met-enkephalin (10^-7M) increased (average 31%) the intracellular calcium level after longer (15 min) treatment. Ionophore A23187 (10^-7M, 10^-6M) was used as a positive control to enhance intracellular calcium level. Thus, δ-opioid agonist DADLE decreased basal intracellular calcium level in MOLT-4 cells after short treatment, while endogenous Met-enkephalin altered intracellular calcium level in a bidirectional way by decreasing and increasing it.     

Fluvastatin inhibits growth and alters the malignant phenotype of the C6 glioma cell line

BackgroundFluvastatin is a member of the family of HMG-CoA reductase inhibitors (statins) extensively used in medical practice. Increasing evidence suggests that fluvastatin may be implicated in suppression of cancer growth and development. The aim of the present study was to investigate the anti-cancer potential of fluvastatin in C6 rat malignant glioma cells.MethodsFirst, the effects of fluvastatin on cell viability (MTT assay), proliferation (BrdU assay), cell morphology, and cytoskeleton were examined. Subsequently, its effect on extracellular signal regulated kinase 1 and 2 (ERK1/2) and c-Jun N-terminal kinase 1 and 2 (JNK 1/2) expression was estimated by Western blot. Finally, the influence of fluvastatin on cell migration and production of MMP-9 and VEGF was determined using a wound-healing assay and ELISA test, respectively.ResultsThe results obtained showed that fluvastatin had a remarkable inhibitory and cytotoxic effect on tumor C6 cells (IC₅₀ = 8.6 μM, 48 h), but did not inhibit the growth of normal neuronal cells. The concentrations from 1 to 10 μM induced marked morphologic alterations typical for apoptosis including shrinkage of cytoplasm, chromatin condensation, and nucleus breakdown.ConclusionThe inhibitory effects of fluvastatin on cell proliferation seemed to be associated with decreased p-ERK1/2 expression, upregulation of p-JNK1/2, and reduction in the MMP-9 and VEGF concentrations in culture media. The high anticancer (antiproliferative, proapoptotic, antiinvasive) activity of fluvastatin and lack of its toxicity against normal cells indicate a potential use of this statin in the treatment of malignant glioma.     

Saikosaponins-b suppresses tumor growth and angiogenesis of hepatocellular carcinoma by regulating VEGF/ERK/HIF-1α signal pathway

OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosaponins(SS)-b on hepatocellular carcinoma(HCC)and its regulation on VEGF/ERK/HIF-1 αsignal pathway.METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro.Pathological change of tumor tissue was observed by HE staining,the microvascular changes were detected by immunohistochemical method.The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane(CAM)model.The effects of SS-b on proliferation,migration and invasion were investigated by MTT assay,scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell(HUVEC)and HepG2 cells in vitro.Vascular endothelial growth factor(VEGF),matrix metalloproteinase-2/9(MMP-2/9),hypoxia-inducible factor-1α(HIF-1α)expression and the phosphorylation of extracellular regulated kinase(ERK)were analyzed using RT-PCR and Westernblot.RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo.The inhibitory rate of tumor was 49.1%,50.7%,66.1%in SS-b 5,10 and 20 mg·kg-1group respectively.HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice.Moreover,SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF.SS-b had an obvious inhibitory effect on cell proliferation,migration and invasion of HUVEC cells and HepG-2 cells.These effects were associated with downregulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1αsignaling in H22 mice and Hep-G2 cells.CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibiting tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.     

Butein suppresses the expression of nuclear factor-kappa B-mediated matrix metalloproteinase-9 and vascular endothelial growth factor in prostate cancer cells

3,4,2′,4′-Tetrahydroxychalcone (butein) has potent anti-inflammatory, anti-cancer and anti-fibrogenic effects. However, little is known about the mechanism by which butein inhibits metastasis and invasion. This study aimed to investigate the effects of butein on the expression of matrix metalloproteinase (MMP-9) and vascular endothelial growth factor (VEGF) in human prostate cancer cells. Butein in vitro resulted in a moderate inhibition of cell proliferation and viability through G₂/M phase arrest. We also analyzed the effect of butein on the activities of nuclear factor-kappa B (NF-κB)-regulated MMP-9 and VEGF since they are prominently involved in the processes of tumor cell invasion and metastasis. Our results in vitro showed that butein attenuates VEGF and MMP-9 activities via the suppression of NF-κB activity. Furthermore, butein repressed the expression of VEGF and MMP-9 induced by treatment with tumor necrosis factor-α and phorbol-12-myristate-13-acetate. Taken together, these data suggest that a blockade of NF-κB activity by butein inhibits invasion and angiogenesis in prostate cancer cells.     

VEGF与MMP-9在卵巢癌组织中的表达及与淋巴结浸润转移的相关性研究

目的探讨VEGF与MMP-9在卵巢癌组织中的表达,进一步探讨二者与淋巴结浸润转移的相关性与作用机制。方法采用免疫组织化学方法检测34例卵巢癌组织中VEGF、MMP-9的表达,并运用统计学方法对二者与卵巢癌的临床病理特征的关系进行对比分析。同期选择正常卵巢组织30例、卵巢良性肿瘤30例作对照。结果 VEGF、MMP-9在卵巢癌组织中阳性表达显著高于正常组织(P<0.05),VEGF、MMP-9的阳性表达与年龄、组织分化类型无相关性(P>0.05);VEGF、MMP-9在有淋巴结转移组织中的表达显著高于无淋巴结转移组织(P<0.05);VEGF和MMP-9在FIGO分期为Ⅲ~Ⅳ的组织中阳性表达显著高于Ⅰ~Ⅱ期组织(P<0.05)。VEGF表达和MMP-9在卵巢组织中表达呈正相关(r=0.625,P<0.05)。结论 VEGF、MMP-9均参与卵巢癌的发生和发展,二者高表达使肿瘤组织转移性和侵袭性显著增加。     

Caveolin-1 is important for nitric oxide-mediated angiogenesis in fibrin gels with human umbilical vein endothelial cells

AIM: The role of caveolin-1 (Cav-1) in angiogenesis remains poorly understood. The endothelial nitric oxide (NO) synthase (eNOS), a caveolin-interacting protein, was demonstrated to play a predominant role in vascular endothelial growth factor (VEGF) -induced angiogenesis. The purpose of our study was to examine the role of Cav-1 and the eNOS complex in NO-mediated angiogenesis. METHODS: Human umbilical vein endothelial cells (HUVEC) were isolated and cultured in 3-D fibrin gels to form capillary-like tubules by VEGF stimulation. The expression of Cav-1 and eNOS was detected by semiquantitative RT-PCR. The HUVEC were treated with antisense oligonucleotides to downregulate Cav-1 expression. Both transduced and non-infected HUVEC were cultured in fibrin gels in the presence or absence of VEGF (20 ng/mL) and NG-nitro-L-arginine methyl ester (L-NAME; 5 mmol/L). NO was measured using a NO assay kit and capillary-like tubules were quantified by tubule formation index using the Image J program. RESULTS: RT-PCR analysis revealed that Cav-1 levels steadily increased in a time-dependent manner and reached their maximum after 5 d of incubation, but there were no obvious changes in eNOS mRNA expression in response to VEGF in the fibrin gel model. VEGF (20 ng/mL) can promote NO production and the formation of capillary-like tubules, and this promoting effect of VEGF was blocked by the addition of L-NAME (5 mmol/L). When transduced HUVEC with the antisense Cav-1 oligonucleotides were plated in the fibrin gels, the capillary-like tubules were significantly fewer than those of the non-infected cells. The capillary-like tubules formation and NO production of transduced HUVEC with the antisense Cav-1 oligonucleotides cultured in fibrin gels showed no responses to the addition of VEGF (20 ng/mL) and L-NAME (5.0 mmol/L). CONCLUSION: NO was a critical angiogenic mediator in this model. Cav-1 was essential for NO-mediated angiogenesis and may be an important target of anti-angiogenesis therapy.