色素上皮衍生因子和血管上皮生长因子在良性前列腺增生组织中的表达及意义

目的:探讨色素上皮衍生因子(PEDF)和血管上皮生长因子(VEGF)在BPH组织中的表达特点和意义.方法:对35例手术切除的BPH组织(BPH组),21例正常前列腺组织(对照组)行免疫组织化学法检测PEDF和VEGF的表达水平和微血管密度(MVD)计数,统计学分析两组标本表达水平的差异以及各因素之间的相关性.结果:BPH组中VEGF和MVD表达明显高于对照组(P＜0.01),PEDF表达明显低于对照组(P＜0.01);MVD与VEGF表达呈显著正相关(P＜0.05),与PEDF表达呈显著负相关(P＜0.05);VEGF与PEDF表达呈显著负相关(P＜0.01).结论:PEDF表达水平降低和VEGF表达水平增加,促进前列腺组织的血管生成作用,导致前列腺组织增生.PEDF和VEGF的不平衡表达可能是BPH组织血管生成的分子基础,是促进BPH发生和发展的因素.     

血管内皮生长相关因子在胰腺癌组织中的表达

目的 观察胰腺癌组织中的血管内皮生长相关因子:色素上皮衍生因子(PEDF)和血管内皮生长因子(VEGF)的表达以及胰腺癌中PEDF和VEGF平衡关系的变化与其预后的关系.方法 用SP免疫组织化学方法检测51例胰腺癌标本,观察PEDF和VEGF在胰腺癌中的表达以及相应的微血管密度(MVD),并和肿瘤血管密度、肿瘤浸润深度、肿瘤大小、血管侵犯、肿瘤病理TNM分期、淋巴结转移、远处转移等临床病理指标作单因素和多因素COX回归分析.结果 PEDF/VEGF与MVD之间明显关系(P<0.01),PEDF/VEGF值大小与肿瘤病理TNM分期(P<0.01)、淋巴结转移(P<0.01)、远处转移(P<0.05)有密切关系.Kaplan-Meier生存曲线分析,51例患者根据PEDF/VEGF值是否大于0.5分为两组,其中PEDF/VEGF≥0.5组生存曲线明显高于PEDF/VEGF<0.5组(log-rank test,P<0.01).PEDF/VEGF比值(风险系数0.390;P<0.05)、淋巴结转移(风险系数3.38;P<0.01)、远处转移(风险系数3.447;P<0.01)是胰腺癌独立的预后指标.结论 PEDF/VEGF比值可较好反映胰腺癌中的血管增生;它与胰腺癌患者的临床分期、淋巴结转移及远处转移相关,是胰腺癌重要的预后因子之一.     

血管内皮生长相关因子在胰腺癌组织中的表达

目的 观察胰腺癌组织中的血管内皮生长相关因子:色素上皮衍生因子(PEDF)和血管内皮生长因子(VEGF)的表达以及胰腺癌中PEDF和VEGF平衡关系的变化与其预后的关系.方法 用SP免疫组织化学方法检测51例胰腺癌标本,观察PEDF和VEGF在胰腺癌中的表达以及相应的微血管密度(MVD),并和肿瘤血管密度、肿瘤浸润深度、肿瘤大小、血管侵犯、肿瘤病理TNM分期、淋巴结转移、远处转移等临床病理指标作单因素和多因素COX回归分析.结果 PEDF/VEGF与MVD之间明显关系(P<0.01),PEDF/VEGF值大小与肿瘤病理TNM分期(P<0.01)、淋巴结转移(P<0.01)、远处转移(P<0.05)有密切关系.Kaplan-Meier生存曲线分析,51例患者根据PEDF/VEGF值是否大于0.5分为两组,其中PEDF/VEGF≥0.5组生存曲线明显高于PEDF/VEGF<0.5组(log-rank test,P<0.01).PEDF/VEGF比值(风险系数0.390;P<0.05)、淋巴结转移(风险系数3.38;P<0.01)、远处转移(风险系数3.447;P<0.01)是胰腺癌独立的预后指标.结论 PEDF/VEGF比值可较好反映胰腺癌中的血管增生;它与胰腺癌患者的临床分期、淋巴结转移及远处转移相关,是胰腺癌重要的预后因子之一.     

色素上皮衍生因子和血管内皮生长因子在胰腺癌中表达

目的探讨色素上皮衍生因子PEDF和血管内皮生长因子VEGF在胰腺癌组织中的表达以及胰腺癌中PEDF和VEGF平衡关系的变化与其预后的关系。方法选择1993年至2005年在中山大学附属第二医院行手术治疗的胰腺癌病人,共51例,用SP免疫组化的方法对胰腺癌标本进行检测,观察两者在胰腺癌中的表达情况及联系,与肿瘤血管密度和肿瘤浸润深度、肿瘤大小、血管侵犯,肿瘤病理TNM分期,淋巴结转移、远处转移等临床病理指标作单因素和多因素COX回归分析。结果本组51例均获得随访,中位随访时间40.3个月（6-156个月）PEDF/VEGF与MVD之间存在显著关系,PEDF/VEGF值大小与肿瘤病理TNM分期,淋巴结转移、远处转移有密切关系。Kaplan-Meier生存曲线分析,51例病人根据PEDF/VEGF值是否大于0.5分为两组,其中PEDF/VEGF≥0.5组生存曲线明显高于PEDF/VEGF〈0.5组（log-ranktest,P=0.0005）。PEDF/VEGF比值（风险系数0.390;P=0.027）、淋巴结转移（风险系数3.38;P=0.005）、远处转移（风险系数3.447;P=0.003）是胰腺癌独立的预后指标。结论PEDF/VEGF比值可较好的反映胰腺癌中的血管增生情况;它与胰腺癌病人的临床分期、淋巴结转移及远处转移相关,系胰腺癌重要的预后因子之一,可以为胰腺癌的抗血管增生治疗提供新的靶点。     

血管内皮生长相关因子在胰腺癌组织中的表达

目的 观察胰腺癌组织中的血管内皮生长相关因子:色素上皮衍生因子(PEDF)和血管内皮生长因子(VEGF)的表达以及胰腺癌中PEDF和VEGF平衡关系的变化与其预后的关系.方法 用SP免疫组织化学方法检测51例胰腺癌标本,观察PEDF和VEGF在胰腺癌中的表达以及相应的微血管密度(MVD),并和肿瘤血管密度、肿瘤浸润深度、肿瘤大小、血管侵犯、肿瘤病理TNM分期、淋巴结转移、远处转移等临床病理指标作单因素和多因素COX回归分析.结果 PEDF/VEGF与MVD之间明显关系(P<0.01),PEDF/VEGF值大小与肿瘤病理TNM分期(P<0.01)、淋巴结转移(P<0.01)、远处转移(P<0.05)有密切关系.Kaplan-Meier生存曲线分析,51例患者根据PEDF/VEGF值是否大于0.5分为两组,其中PEDF/VEGF≥0.5组生存曲线明显高于PEDF/VEGF<0.5组(log-rank test,P<0.01).PEDF/VEGF比值(风险系数0.390;P<0.05)、淋巴结转移(风险系数3.38;P<0.01)、远处转移(风险系数3.447;P<0.01)是胰腺癌独立的预后指标.结论 PEDF/VEGF比值可较好反映胰腺癌中的血管增生;它与胰腺癌患者的临床分期、淋巴结转移及远处转移相关,是胰腺癌重要的预后因子之一.     

Adeno-associated virus vector-mediated delivery of pigment epithelium-derived factor restricts neuroblastoma angiogenesis and growth

Purpose

The purpose of this study was to evaluate the ability of adeno-associated virus (AAV) vector-mediated delivery of pigment epithelium-derived factor (PEDF) to inhibit neuroblastoma (NB) xenograft growth. Pigment epithelium-derived factor was chosen for this study because, in addition to being a potent inhibitor of angiogenesis, it is capable of inducing neuronal differentiation.

Methods

Cohorts of mice received either recombinant AAV encoding human PEDF (rAAV-hPEDF) at a range of doses or control vector via tail vein. Subsequent hPEDF expression was measured by enzyme-linked immunoassay. After 6 weeks, the mice were given human NB cells by retroperitoneal injection and then killed 5 weeks later. Tumor weight, microvessel density, tumor differentiation, apoptosis, and levels of intratumoral vascular endothelial growth factor (VEGF) expression were determined at that time. In subsequent cohorts of mice, AAV-mediated murine PEDF expression was tested against both human NB xenografts and murine tumors.

Results

In a series of in vitro studies, PEDF was shown to inhibit endothelial cell activation and to stimulate differentiation of NB cell lines. After tail vein injection of rAAV-hPEDF, stable transgene expression was generated and correlated with levels of vector administration. Human NB xenograft growth was restricted by hPEDF in a dose-dependent fashion. Intratumoral VEGF expression and microvessel density were decreased, and tumor cell apoptosis was increased in PEDF-treated mice.

Conclusions

Treatment with PEDF had a significant impact on NB growth in mice when delivered continuously using a gene therapy-mediated approach. The activity of PEDF appears to be mediated in part by inhibition of tumor-induced angiogenesis through down-regulation of tumor-elaborated VEGF, with subsequent intratumoral apoptosis. Furthermore, hPEDF was able to induce NB differentiation in vitro and in vivo. In addition, antitumor efficacy was seen when mouse PEDF was used to treat syngeneic murine tumors. In our murine models, gene therapy-mediated delivery of PEDF appears promising for the treatment of neuroblastoma.     

活体转染肝细胞生长因子基因对小鼠急性肝功能衰竭的治疗作用

目的 研究肝细胞生长因子（HGF）基因治疗对小鼠急性肝衰模型的治疗作用。方法 构建人肝细胞生长因子（hHGF）表达载体，利用脂质体介导法在活体内转染肝细胞生长因子基因，利用荧光显微镜检测肝组织内绿色荧光蛋白（GFP）表达了解目的基因的表达，用酶联免疫吸附试验（ELISA）法检测血清中人肝细胞生长因子的含量，通过观察小鼠急性肝衰模型的生存率、肝功能变化、肝组织病理改变来检测肝细胞生长因子基因对急性肝衰的治疗作用，通过检测肝组织中PC—NA指数的变化了解肝脏的增殖能力的变化。结果 荧光显微镜下可见肝组织内有绿色荧光蛋白的表达，转染人肝细胞生长因子基因后在血清中可检测到hHGF的表达，而且可持续1周以上，与对照组相比。转染hHGF基因组的生存率明显提高（40．0％vs11．5％，P〈0．05），血清ALT、TBi明显降低，肝组织中PCNA指数也明显升高。结论 活体转染肝细胞生长因子基因可获得表达，而且肝细胞生长因子基因对急性肝衰小鼠有治疗作用。     

In vivo electroporation enhances plasmid-based gene transfer of basic fibroblast growth factor for the treatment of ischemic limb

BACKGROUND: Angiogenic therapy for ischemic tissues using angiogenic growth factors has been reported on an experimental and a clinical level. Electroporation enhances the efficiency of plasmid-based gene transfer in a variety of tissues. The purpose of this study was to evaluate the angiogenic effects of plasmid-based gene transfer using basic fibroblast growth factor (bFGF) in combination with electroporation. MATERIALS AND METHODS: The transfection efficiency of in vivo electroporation in rabbit skeletal muscles was evaluated using pCAccluc+ encoding luciferase. To evaluate the angiogenic effects of bFGF gene in ischemic limb, we constructed a plasmid, pCAcchbFGFcs23, containing human bFGF cDNA fused with the secretory signal sequence of interleukin (IL)-2. Then, 500 microg of pCAcchbFGFcs23 or pCAZ3 (control plasmid) was injected into the ischemic thigh muscles in a rabbit model of hind limb ischemia with in vivo electroporation (bFGF-E(+) group and LacZ-E(+) group). Other sets of animals were injected with pCAcchbFGFcs23 (bFGF-E(-) group) or pCAZ3 (LacZ-E(-) group) without electroporation. Then 28 days later, calf blood pressure ratio, angiographic score, in vivo blood flow, and capillary density in the ischemic limb were measured. RESULTS: Gene transfer efficiency increased markedly with the increase in voltage up to 100 V. Regarding angiogenic responses, calf blood pressure ratio, in vivo blood flow, and capillary density only in the bFGF-E(+) group were significantly higher than those in LacZ-E(-) group. Angiographic scores in the bFGF-E(+) and bFGF-E(-) groups were significantly higher than that in the LacZ-E(-) group. CONCLUSION: These data suggest that in vivo electroporation enhances bFGF gene transfer for the treatment of ischemic limb muscles.     

Retroviral endostatin gene transfer inhibits growth of human lung cancer in a murine orthotopic xenotransplant model

Background and aims The administration of endostatin, a potent anti-angiogenic agent, will be required for extended periods of time as a cancer treatment. The aim of the present study was to induce endogenous endostatin secretion in a continuous fashion, based on retroviral gene transfer. The tumor response was evaluated in an orthotopic murine tumor model of human lung cancer.Materials and methods Human non-small-cell lung cancer cells (KNS 62) were retrovirally transduced with the human endostatin gene. An orthotopic murine xenotransplant model was used to investigate tumor growth, metastases and survival. After 4 weeks of subcutaneous growth, endostatin expression was measured by immunoblot analysis in tumor lysates.Results The growth of the subcutaneous tumors was significantly delayed, and orthotopic tumor growth and pleural metastases were significantly reduced in endostatin-transduced KNS 62 tumors. Prolongation of survival subsequent to orthotopic tumor induction was demonstrated. Strong endostatin expression was found in subcutaneous tumors after 4 weeks.Conclusion Retroviral transduction of the human endostatin gene is capable of achieving long-term endostatin expression. Endostatin transduction provides significant anti-tumor effects with regard to local tumor growth, metastases and survival.     

Neutralizing vascular endothelial growth factor activity inhibits thyroid cancer growth in vivo

BACKGROUND: Without angiogenesis, tumor growth is limited to a few millimeters, the limit of diffusion. Vascular endothelial growth factor (VEGF) is an endothelial-specific mitogen and a major regulator of angiogenesis. METHODS: To investigate the relationship between VEGF and thyroid tumor angiogenesis, we xenografted human dermal matrix inoculated with FTC-133 cells into nude mice or directly injected FTC-133 cells subcutaneously. To block the function of VEGF, the neutralizing anti-VEGF monoclonal antibody A.4.6.1 (mAb A.4.6.1) was injected intraperitoneally twice weekly. As control, an antibody of the same isotype (Ab 5B6) or phosphate buffer saline solution (PBS) was used. To evaluate the dermal matrix as a model for angiogenesis studies, recombinant human VEGF was inoculated into the dermal matrix pocket and xenografted into mice. RESULTS: In the dermal matrix angiogenesis model, the number of blood vessels paralleled the concentration of recombinant human VEGF and was highest at 100 ng/mL. Mice that were treated with the mAb A4.6.1 developed fewer blood vessels (mean, 6.6 per HPF) than control mice (18 per HPF in Ab 5B6 and 22 per HPF in PBS; P <.01). Tumors from mice that were treated with mAb A.4.6.1 were much smaller (mean +/- SD, 0.09 +/- 0.02 gm) at 5 weeks, compared with the tumors treated with Ab 5B6 (5.38 +/- 1.15 gm) or PBS (4.0 +/- 0.72 gm; P <.001). CONCLUSIONS: VEGF is produced by the follicular thyroid cancer cell line and stimulates angiogenesis and growth of thyroid cancer. This stimulation can be blocked by mAb A.4.6.1.     

In vivo hepatocyte growth factor gene transfer to bladder smooth muscle after bladder outlet obstruction in the rat: a morphometric analysis

PURPOSE: We determined whether hepatocyte growth factor gene transfer after partial bladder outlet obstruction would prove effective for decreasing transforming growth factor-beta expression and consequently decreasing collagen deposition in partially obstructed rat bladders. MATERIALS AND METHODS: Ten-week-old male Sprague-Dawley rats were divided into 3 groups of 10 each, including group 1--sham operation, group 2--bladder outlet obstruction for 4 weeks and group 3--hepatocyte growth factor gene transfer after bladder outlet obstruction. Two weeks after the onset of bladder outlet obstruction in group 3 hepatocyte growth factor-liposome complex (50 microg human hepatocyte growth factor cDNA) was injected into the smooth muscle of the rats. RESULTS: We noted no difference between groups 2 and 3 with regard to the ratio of bladder weight to body weight. The ratio in groups 2 and 3 was significantly higher than in group 1 (p = 0.043). The mean percent of collagen area +/- SE was 36.32% +/- 1.83%, 27.90% +/- 2.66% and 8.97% +/- 3.35% in groups 1 to 3, respectively (p <0.05). Relative hepatocyte growth factor and c-met mRNA and protein expression were higher in group 3 than in groups 1 and 2. However, the expression of transforming growth factor-beta1 mRNA and protein was higher in group 2 than in groups 1 and 3. CONCLUSIONS: These findings may imply a possible novel therapeutic strategy against bladder dysfunction arising in patients with bladder outlet obstruction.     

Decreased expression of pigment epithelium-derived factor is involved in the pathogenesis of diabetic nephropathy

Pigment epithelium-derived factor (PEDF) is a potent angiogenic inhibitor. Previous studies have shown that decreased ocular levels of PEDF are associated with diabetic retinopathy. However, the implication of PEDF expression in diabetic nephropathy has not been revealed. In the present study, we demonstrated for the first time that the expression of PEDF was decreased at both the mRNA and protein levels in the kidney of diabetic rats, whereas transforming growth factor-beta (TGF-beta) and fibronectin levels were increased in the same diabetic kidneys. As shown by immunohistochemistry, the decrease of PEDF expression occurs primarily in the glomeruli. In vitro studies showed that high concentrations of glucose significantly decreased PEDF secretion in primary human glomerular mesangial cells (HMCs), suggesting that hyperglycemia is a direct cause of the PEDF decrease in the kidney. Toward the function of PEDF, we showed that PEDF blocked the high-glucose-induced overexpression of TGF-beta, a major pathogenic factor in diabetic nephropathy, and fibronectin in primary HMCs, suggesting that PEDF may function as an endogenous inhibitor of TGF-beta expression and fibronectin production in glomeruli. Therefore, decreased expression of PEDF in diabetic kidneys may contribute to extracellular matrix overproduction and the development of diabetic nephropathy.     

Loss of the antiangiogenic pigment epithelium-derived factor in patients with angiogenic eye disease.

Retinal neovascularization characterizes proliferative diabetic retinopathy (PDR). Pigment epithelium-derived factor (PEDF) has been shown to be a major antiangiogenic growth factor in the mammalian eye. PEDF expression is suppressed by hypoxia, and changes in PEDF have been correlated to the development of retinal neovascularization in animal models of hypoxic eye disease. However, whether this concept of a reduced angiogenesis inhibitor holds true in humans is as yet unclear. In this study, we analyzed the in vivo regulation of PEDF in patients with and without hypoxic eye disease. We used immunoblots to measure PEDF in ocular fluids obtained from 64 nondiabetic and diabetic patients. In addition, immunohistochemistry of PEDF was carried out in specimens of normal human retinas and retinas with various degrees of diabetic retinopathy. The PEDF concentrations in patients with PDR (P < 0.001) or extensive nondiabetic retinal neovascularization caused by retinal-vein occlusion (P < 0.001) were lower than in control patients. Levels of PEDF were replenished in PDR patients with previous retinal scatter photocoagulation compared with PDR patients without previous photocoagulation (P = 0.01). Immunohistochemistry revealed an interstitial staining pattern as expected for a secreted protein, with an intense staining in retinas of patients without proliferative eye disease. However, in patients with PDR, little or no staining was detectable. Our data strongly support the concept that retinal angiogenesis is induced by loss of the major angiogenesis inhibitor in the eye, PEDF, in combination with an increased expression of angiogenic growth factors such as vascular endothelial growth factor. Our findings suggest that substitution of angiogenesis inhibitors may be an effective approach in the treatment of PDR.     

Salutary effect of pigment epithelium-derived factor in diabetic nephropathy: evidence for antifibrogenic activities

Diabetic nephropathy is a major complication of diabetes and a leading cause of end-stage renal diseases in the U.S. Pigment epithelium-derived factor (PEDF) is a potent angiogenic inhibitor that has been extensively studied in diabetic retinopathy. Recently, we reported that PEDF is expressed at high levels in normal kidneys and that PEDF levels are decreased in kidneys of streptozotocin (STZ)-induced diabetic rats. In the present study, we injected STZ-diabetic rats with an adenovirus expressing PEDF (Ad-PEDF) to evaluate its effects in diabetes. The results showed that increased expression of PEDF in the kidney in response to Ad-PEDF delivery significantly alleviated microalbuminuria in early stages of diabetes. Administration of Ad-PEDF was found to prevent the overexpression of two major fibrogenic factors, transforming growth factor-beta (TGF-beta)1 and connective tissue growth factor (CTGF), and to significantly reduce the production of an extracellular matrix (ECM) protein in the diabetic kidney. Moreover, PEDF upregulated metalloproteinase-2 expression in diabetic kidney, which is responsible for ECM degradation. In cultured human mesangial cells, PEDF significantly inhibited the overexpression of TGF-beta1 and fibronectin induced by angiotensin II. PEDF also blocked the fibronectin production induced by TGF-beta1 through inhibition of Smad3 activation. These findings suggest that PEDF functions as an endogenous anti-TGF-beta and antifibrogenic factor in the kidney. A therapeutic potential of PEDF in diabetic nephropathy is supported by its downregulation in diabetes; its prevention of the overexpression of TGF-beta, CTGF, and ECM proteins in diabetic kidney; and its amelioration of proteinuria in diabetic rats following Ad-PEDF injection.     

基质金属蛋白酶-9和色素上皮衍生因子在胰腺癌侵袭转移中的作用

目的 观察基质金属蛋白酶-9(MMP-9)和色素上皮衍生因子(PEDF)基因表达在胰腺癌(PC)侵袭转移中的作用。方法 应用免疫组织化学法检测35例胰腺癌标本中癌细胞的MMP-9和PEDF的表达,统计分析两种分子标记物与胰腺癌细胞MMP-9和PEDF的表达。结果 胰腺癌组中MMP-9和PEDF表达率分别为71.4％和34.3％,MMP-9高表达率和PEDF低表达率与浸润范围、淋巴转移和远处转移呈正相关(P＜0.05)。MMP-9与PEDF表达呈负相关(P＜0.01)。结论 MMP-9在胰腺癌侵袭转移中起促进作用,PEDF可抑制肿瘤生长,PEDF低表达可能通过上调MMP-9的表达,从而促进胰腺癌的侵袭转移。