非小细胞肺癌bFGF表达与微血管密度的意义

目的：探讨非小细胞肺癌（ＮＳＣＬＣ）组织中碱性成纤维细胞生长因子（ｂＦＧＦ）表达和微血管密度（ＭＶＤ）的意义。方法：采用免疫组化ＡＢＣ法检测７８例ＮＳＣＬＣ标本中ｂＦＧＦ表达和ＭＶＤ，并根据病理类型、组织学分化程度、有无区域淋巴结转移以及预后进行分析。结果：在ＮＳＣＬＣ中腺癌的ｂＦＧＦ表达阳性率和ＭＶＤ值显著高于鳞癌（Ｐ〈０．０５）；所有ＮＳＣＬＣ病例中，ｂＦＧＦ表达阳性率与肿瘤组织学分化程度有关     

微血管定量和血管内皮生长因子表达在肠型胃癌中的意义

为研究血管形成和血管形成因子表达在肠型胃癌和弥漫型胃癌中的作用，应用抗因子Ⅷ相关抗原抗体、抗ＶＥＧＦ抗体及抗ｂＦＧＦ抗体的免疫组化ＬＳＡＢ法，分别对６３例肠型胃癌和４５例弥漫型胃癌中的微血管数量（ＭＶＣ）、ＶＥＧＦ和ｂＦＧＦ表达进行研究。ＭＶＣ和ＶＥＧＦ及ｂＦＧＦ表达在肠型胃癌明显高于弥漫型胃癌（Ｐ分别＜０．０５，０．００１和０．０１），同样，ＭＶＣ和ＶＥＧＦ表达在肝转移患者明显高于腹膜转移者（Ｐ分别＝０．００１和＜０．０１）；在肠型胃癌中，ＭＶＣ与ＶＥＧＦ表达明显相关（Ｐ＝０．０２），ＭＶＣ和ＶＥＧＦ表达随ＴＮＭ分期增加而增加，而与弥漫型胃癌无关。两型胃癌中的ｂＦＧＦ表达均与ＭＶＣ无关。本研究结果表明肠型胃癌的转移形式为血管依赖性，ＶＥＧＦ可能是诱导肠型胃癌血管形成的一个重要因子。     

膀胱癌中bFGF和血管形成的研究

目的：探讨碱性成纤维细胞生长因子（ｂＦＧＦ）和血管形成与膀胱癌的发病及生物学行为之间关系。方法：采用免疫组化方法检测７２例膀胱癌组织和２１例正常膀胱组织中ｂＦＧＦ和第ＶＩＩＩ因子相关抗原表达。结果：膀胱癌组织中ｂＦＧＦ表达和微血管密度（ＭＶＤ）均显高于正常膀胱胱组织，而且两与肿瘤病理分级分期均密切相关。肿瘤复发的ｂＦＧＦ表达和ＭＶＤ均显高于复发，ｂＦＧＦ强阳性表达的ＭＶＤ值显高于阴性及弱阳性表达，结论：ｂＦＧＦ通过促进血管形成而在膀胱癌的发生发展过程中起着重要的作用，而且ｂＦＧＦ和ＭＶＤ可作为膀胱癌生物行为和预后的评估指标。     

亚硝胺诱发大鼠肝癌表达转化生长因子β与肥大细胞浸润的关系

目的 探讨肝癌转化因子β（ｔｒａｎｓｆｏｒｍｉｎｇ ｇｒｏｗｔｈ ｆａｃｔｏｒ β,ＴＧＦβ）表达及与癌周肥大细胞（ｍａｓｔ ｃｅｌｌ,ＭＣ）数量的相关性。方法 二乙基硝胺诱发大鼠肝癌模型,应用光镜、电镜、免疫组织化学及细胞图像分析技术观察肝癌细胞ＴＧＦβ表达与癌周ＭＣ数量变化之间的关系。结果 肝癌细胞胞浆中ＴＧＦβ呈阳性表达,不同组别的大鼠肝癌ＴＧＦβ表达强度在各癌巢间显示明显的差异,ＴＧＦβ表达强弱与癌周ＭＣ数量变化呈平行一致关系（Ｐ〈０．０１）。结论 肝癌细胞通过释放细胞因子ＴＧＦβ并以旁分泌方式影响癌周ＭＣ的数量,ＴＧＦβ是影响癌周ＭＣ数量变化的相关因素之一。     

碱性成纤维细胞生长因子在膀胱移行细胞癌的表达及意义

目的：研究碱性成纤维细胞生长因子（ｂＦＧＦ）在膀胱移行细胞癌（ＢＴＣＣ）的表达及其意义。方法：应用免疫组化ＡＢＣ法检测３３例ＢＴＣＣ及１２例正常膀胱组织中ｂＦＧＦ抗原。结果：ｂＦＧＦ在正常移行细胞呈阴性表达，在正常组织和肿瘤组织的血管内皮细胞、平滑肌细胞均呈弱性表达，肿瘤细胞呈不同程度的阳性表达且过表达率在不同病理分级临床分期间有显著性差异（Ｐ〈０．０５），在单发与多发、初发与复发组间无显著性差异     

喉粘膜癌前病变和癌变中VEGF、bFGF和KGF的表达分析

目的　探讨血管生成因子在喉癌发生发展中的作用。方法　本文用免疫组织化学和原位杂交方法,观察ＶＥＧＦ、ｂＦＧＦ和ＫＧＦ在喉粘膜不典型增生（ＤＹＳ）及鳞癌（ＳＣＣ）中的表达和分布情况。结果　ＶＥＧＦ、ｂＦＧＦｍ ＲＮＡ、ＫＧＦ ｍ ＲＮＡ 在ＳＣＣ 中的表达较ＤＹＳ有所增强,其中ＶＥＧＦ不仅在上皮细胞中表达,间质中部分血管内皮细胞及炎性细胞亦表达ＶＥＧＦ;而ｂＦＧＦ、ＫＧＦ的转录仅在ＤＹＳ和ＳＣＣ的间质中出现,上皮细胞未见阳性表达,ｂＦＧＦ、ＫＧＦ阳性反应主要位于血管内皮细胞和成纤维细胞。结论　在喉癌中ＶＥＧＦ、ｂＦＧＦ、ＫＧＦ可能分别通过自分泌或旁分泌方式促进间质中新生血管和间质的形成,直接或间接地影响肿瘤细胞的生长。     

慢性粒细胞白血病患者血小板膜糖蛋白Ⅳ再分布和胞内凝血酶敏感蛋白的释放

目的：探讨慢性粒细胞白血病（ＣＭＬ）患者血小板膜糖蛋白Ⅳ（ＧＰⅣ）再分布和胞内凝血酶敏感蛋白（ＴＳＰ）释放及血浆中β－血小板球蛋白（β－ＴＧ）和血小板第４因子（ＰＦ４）对其的影响作用。方法：应用１２５Ｉ分别标记血小板ＧＰⅠｂ、ＧＰⅡｂ／Ⅲａ、ＧＰⅣ和ＴＳＰ单克隆抗体并结合放射免疫法检测ＣＭＬ患者血小板膜上相应的结合位点，同时观察β－ＴＧ和ＰＦ４的抑制作用。结果：（１）ＣＭＬ患者未活化和诱导活化（凝血酶１Ｕ／ｍｌ）血小板膜ＧＰⅣ分布和再分布分别为３６０８０±１７０１０、４４３２０±３２３１０抗体结合分子数／血小板，明显高于相应对照组（Ｐ＜０．０１），而血小板膜ＧＰⅠｂ和ＧＰⅡｂ／Ⅲａ则无异常分布；（２）ＣＭＬ患者血小板胞内ＴＳＰ释放并未伴随血小板膜ＧＰⅣ再分布而发生相应的增多；（３）ＣＭＬ患者血浆中β－ＴＧ和ＰＦ４含量增高（Ｐ＜０．０５），体外实验发现β－ＴＧ和ＰＦ４明显地抑制血小板胞内ＴＳＰ释放。结论：ＣＭＬ患者血小板膜ＧＰⅣ再分布增多，可能成为判断血小板功能异常指标之一；而血小板胞内ＴＳＰ释放障碍或结合下降，则受血浆中β－ＴＧ和ＰＦ４的影响。     

碱性成纤维细胞生长因子（BFGF）诱变性研究

本文应用活体小鼠骨髓细胞染色体畸变（ＣＡ）试验及骨髓嗜多染红细胞（ＰＣＥ）微核（ＭＮ）试验，对碱性成纤维细胞生长因子（ＢＦＧＦ）的诱变活性进行为期ｌＷＫ、２ＷＫ及４ＷＫ的实验研究。结果表明：经ＢＦＧＦ处理的实验各组微核率与正常对照组比较无显著性差异（Ｐ＞０．０５），但是，染色体畸变试验结果揭示，其与对照组比较显示出一定的诱变活性。ＣＡ及ＭＮ两种方法测定结果均无剂量效应关系。     

碱性性纤维细胞生长因子（BFGF）诱变性研究

本文应用活体小鼠骨髓细胞染色体畸变（ＣＡ）试验及骨髓嗜多染红细胞（ＰＣＥ）微核（ＭＮ）试验，对碱性成纤维细胞生长因子（ＢＦＧＦ）的诱变活性进行为期１ＷＫ、２ＷＫ及４保实验研究。结果表明：经ＢＦＧＦ处理的实验各组微核率与正常对照组比较无显著性差异，但是，染色体畸变试验结果提示，其与对照组比较显示出一定的诱变活性。ＣＡ及ＭＮ两种方法测定结果均无剂量效应关系。     

膀胱移行细胞癌中碱性成纤维细胞生长因子表达的意义

目的 探讨膀胱移行细胞癌中碱性成纤维细胞生长因子（ｂＦＧＦ）的表达及其与临床病理指标的关系。方法 采用免疫组织化学方法，对８例正常膀胱组织及６２例原发性膀胱移行细胞癌中的ｂＦＧＦ进行检测。结果 膀胱移行细胞癌组织中ｂＦＧＦ阳性表达率为４６．８％。低分化癌和浸润性癌中的ｂＦＧＦ阳性表达率明显高于高分化癌和浅表性癌（Ｐ〈０．０５），复发者的阳性表达率明显高于未复发者（Ｐ〈０．０５），ｂＦＧＦ阳性表达者     

碱性成纤维细胞生长因子小分子干扰RNA对胶质瘤细胞株U251增殖与凋亡的影响

背景与目的:碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)是成纤维细胞生长因子(fibroblast growth factors,FGFs)家族中的重要一员,bFGF是多功能细胞因子,参与创伤愈合、组织修复、未成熟神经细胞的增殖和分化过程.本课题组前期对bFGF在胶质瘤中的表达做过研究,证实bFGF在胶质瘤细胞中过表达,并刺激胶质瘤细胞的增殖和新生血管生成.本研究检测bFGF小分子干扰RNA对胶质瘤细胞的增殖和凋亡的影响.方法:用化学法合成四条针对bFGF的siRNA和一条阴性对照siRNA,并用脂质体法转染胶质瘤细胞系U251;以Opti-MEM I无血清培养基培养的U251细胞作为正常对照.通过RT-PCR和免疫组化的方法检测bFGF的表达,并用流式细胞术、MTT法检测转染后U251细胞的凋亡和增殖活性的变化.结果:转染48 h后,正常对照、阴性对照、siRNA-1、siRNA-2、siRNA-3和siRNA-4组U251细胞中的bFGF mRNA水平分别为0.95 0.02、0.95±0.02、0.85 0.02、0.76±0.04、0.65±0.04和0.56±0.03;转染72 h后,分别为0.95±0.04、0.89±0.05、0.81±0.05、0.80±0.05、0.77±0.04和0.53±0.05.四条bFGF siRNA中,siRNA-4作用最显著.转染48 h后,siRNA-4和阴性对照组细胞增殖抑制率分别为(66.4±1.2)%和(56.3±1.4)%;转染72 h后,分别为(40.0±2.6)%和(14.7±0.6)%,siRNA-4与阴性对照组相比差异具有统计学意义(P<0.05).结论:理想的bFGF小分子干扰RNA 能够抑制该基因的表达并抑制胶质瘤细胞的增殖活性.     

Production of Basic Fibroblast Growth Factor-like Factor by Cultured Human Cholangiocellular Carcinoma Cells

An extract of cultured human cholangiocellular carcinoma cells (HuCC-T1) was found to contain high mitogenic activity for BALB/c3T3 cells. The growth factor eliciting most of the mitogenic activity was purified and concluded to be identical with basic fibroblast growth factor (bFGF)-like factor on the basis of its molecular weight and heparin-Sepharose elution profile, and the results of immunoblotting and radioimmunoassay. HuCC-T1 cells also secreted bFGF-like factor into serum-free medium. A combination of insulin and transferrin or bovine serum albumin stimulated the growth of HuCC-T1 cells in serum-free medium. However, bFGF did not stimulate their growth in the presence and absence of these supplements. Neutralizing monoclonal antibody against bFGF did not inhibit growth. These results indicate that bFGF-like factor is not a growth factor for this cell line.     

A vaccine targeting basic fibroblast growth factor elicits a protective immune response against murine melanoma

Tumor growth and metastasis are closely related to angiogenesis. Basic fibroblast growth factor(bFGF) is an angiogenic factor, and up-regulated expression of bFGF plays a crucial role in the development and metastasis of melanoma. Therefore, in this study, we sought to achieve antitumor activity by immunity targeting bFGF which would inhibit tumor angiogenesis and simultaneously induce bFGF specific cytotoxic T lymphocytes to kill melanoma cells. A human bFGF protein was used as exogenous antigen, coupled with a saponin-liposome adjuvant formulation to enhance CTL response. The results showed that the immunity induced strong immune response and produced prominent anti-cancer activities. CD31 immunohistochemistry and alginate-encapsulated tumor cell assay displayed that tumor angiogenesis was effectively inhibited. Further, the higher production of IFN-γ and cytotoxic T lymphocyte killing assay suggested that the anti-cancer activities may mainly depend on cellular immune response, which could cause the inhibition of tumor angiogenesis and specific killing of tumor cells by bFGF-specific cytotoxic T lymphocytes. We concluded that immunotherapy targeting bFGF may be a prominent strategy for melanoma, and that the adjuvant formulation of saponin-liposome is very desirable in enhancing cytotoxic T lymphocytes response.     

Fibroblast growth factor-binding protein expression changes with disease progression in clinical and experimental human squamous epithelium

Basic fibroblast growth factor (bFGF) is synthesized by a wide variety of normal and malignant cells. However, bFGF cannot exert its effects unless it gets outside of the cell. Since it lacks a signal sequence to direct secretion, the method by which cells release it remains unclear. A 17 kDa secreted binding protein for bFGF (FGF-BP, HBp-17) is expressed at high levels in squamous cell carcinoma (SCC) and transformed keratinocytes and may act as a chaperone to transport bFGF outside of the cell. In our study, FGF-BP mRNA expression in normal keratinocytes was higher than in 5/5 SCCs. Using a new monoclonal antibody, we demonstrate that FGF-BP can dimerize. Immunoassays demonstrate that normal keratinocytes have a higher level of FGF-BP than SCCs. In normal human squamous epithelium, we observed diffuse, moderate to intense cytoplasmic and membranous expression of FGF-BP. Expression decreased and became focal with disease progression to invasive cancer. Injection of immortalized but non-tumorigenic HaCaT cells transduced with FGF-BP into normal human skin xenografts failed to result in tumors. Transfection of FGF-BP into the SCCs Det 562 and FaDu did not promote tumor growth more than controls, and peri-tumoral microvessel density was lower in FGF-BP-transfected than in control tumors. Taken together, these data suggest that FGF-BP expression in squamous epithelium does not play an important role in progression to invasive carcinoma.     

Angiogenesis and mast cells in non-Hodgkin's lymphoma: a strong correlation in angioimmunoblastic T-cell lymphoma.

Mast cells are likely to play a role in angiogenesis under pathological conditions. Solid tumor growth is dependent on angiogenesis, but the influence of mast cells on angiogenesis in non-Hodgkin's lymphoma, (NHL) is not clear. We investigated mast cell number and vessel count in 61 cases of NHL. We also evaluated expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), both important cytokines for angiogenesis. The number of mast cells was greater in T-cell lymphomas than in B-cell lymphomas. Of the T-cell lymphomas, the greatest number of mast cells was observed in the angioimmunoblastic T-cell lymphoma (AIL). In all NHLs, significant correlation was found between vessel count and the number of mast cells (p < 0.0001) and between vessel count and the number of VEGF-expressing cells (p < 0.05) but not between vessel count and bFGF-expressing cells. Strong correlation was detected between the number of mast cells and the number of VEGF-expressing cells (p < 0.0001) in all NHLs. Double fluorescence staining of VEGF mRNA and mast cell tryptase revealed that mast cells expressed VEGF mRNA. Our data suggest that mast cells play a very important role in angiogenesis by expressing VEGF in NHL, especially in AIL.     

Upregulation of basic fibroblast growth factor in breast carcinoma and its relationship to vascular density, oestrogen receptor, epidermal growth factor receptor and survival

Background: Angiogenesis, the process whereby endothelial cells divide and migrate to form new blood capillaries, has been assessed in tumours by measuring microvessel density. High microvessel density is a significant adverse prognostic factor in breast cancer. The angiogenic factor, basic fibroblast growth factor (bFGF), has been associated with tumourigenesis and metastasis in several human cancers. There are few quantitative studies of bFGF expression in normal tissues compared to cancer.Patients and methods: We have measured bFGF levels in 149 human primary breast carcinomas and assessed the findings in relation to microvessel density, oestrogen receptor (ER) and epidermal growth factor receptor (EGFR).Basic FGF levels were measured by ELISA. Western blotting and immunohistochemistry were carreid out to confirm the presence of bFGF.Results: Levels of bFGF were more than 10-fold higher in tumour cytosols compared to reduction mammoplasty tissue and 3-fold compared to non neoplastic cytosols from the same breast as the tumour (P < 0.0001). Immunohistochemistry showed bFGF protein was localised exclusively in the stroma whereas no bFGF staining was observed in the epithelial cells. High bFGF levels were significantly related to high ER (P = 0.01). Similarly, high bFGF levels were significantly related to low grade (P = 0.046) and to small tumour size (P = 0.04). No significant relationship was observed between bFGF and microvessel count, EGFR or age. In univariate analysis and in a Cox proportional hazard model bFGF did not reach significance for overall or relapse free survival.Conclusions: Our results show that although bFGF is elevated in breast carcinomas compared to normal breast tissue it is not related to microvessel density and it is not an independent predictor of survival in breast cancer patients. Basic FGF may be one of multiple factors that synergise with other growth factors such as VEGF to enhance angiogenesis.     

脑胶质瘤bFGF蛋白表达与增殖活性的研究

 目的　探讨胶质瘤中碱性成纤维生长因子 (basicfibroblastgrowthfactor,bFGF)蛋白与增殖细胞核抗原 (proliferativecellnuclearantigen ,PCNA)蛋白在胶质瘤中的表达及相互关系。 方法　根据WHO标准进行分类 ,采用免疫组化方法 ,对 35例胶质瘤和 7例正常脑组织石蜡标本中bFGF与PCNA蛋白的表达水平进行检测 ,并比较二者的相关性。结果　bFGF和PCNA在高级别胶质瘤中表达较多 ,在低级别胶质瘤中表达较少 ,在正常脑组织中几乎不表达 ,两两比较差异有显著性 (P <0 .0 1) ,且二者呈显著正相关 (r=0 .74 ,P <0 .0 1)。结论　人脑胶质瘤bFGF蛋白的阳性表达与细胞增殖活性和组织学分级密切相关 ,它可能参与了人脑胶质瘤细胞的恶性转化。