EGF、EGF－R在卵巢浆液性良、恶性及交界性肿瘤中的表达

应用免疫组织化学ＡＢＣ方法，检测５２例卵巢浆液性良、恶性及交界性肿瘤中ＥＧＦ、ＥＧＦ－Ｒ的表达。结果显示：ＥＧＦ、ＥＧＦ－Ｒ在卵巢浆液性囊腺瘤中分别为５０％、３７％，交界性病变中分别为６２．５％、１００％。其中ＥＧＦ－Ｒ检测良恶性及交界性病变阳性表达率有显著性差异，Ｐ＜０．０５，提示ＥＧＦ、ＥＧＦ－Ｒ在卵巢发生发展中起不同调节作用，有利于病理的诊断和临床估计预后。     

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。     

转化生长因子β及其II受体在胶质瘤中的表达

目的：探讨转化生长因子β１ ,２（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈ ｆａｃｔｏｒβ１ ,２ ,ＴＧＦβ１ ,２） 及其ＩＩ 受体（ＴＧＦβＴｙｐｅ ⅡＲｅｃｅｐｔｏｒ ,ＴＧＦβＲ Ⅱ） 在胶质瘤表达与其恶性度关系和发生发展关系。方法： 用ＬＳＡＢ 免疫组化法检测星形细胞瘤４１例,其中Ｉ～ＩＩ级１６ 例,ＩＩＩ～ＩＶ 级２５ 例和正常对照１１ 例的ＴＧＦβ１ ,２ 及其ＲＩＩ 表达。结果：半定量示ＴＧＦβ１ ,２ 在正常脑组织、ＩＩＩ 级和ＩＩＩＩＶ 级胶质瘤三者的表达呈现从弱到强的正相关关系,有显著差别（ Ｐ＜ ０－０５） ;ＴＧＦβＲＩＩ在三者表达呈现从强到弱的负相关关系,有显著差别（ Ｐ＜ ０－０１） 。结论： 胶质瘤ＴＧＦβ１ ,２ 从弱到强表达和ＴＧＦβＲＩＩ从强到弱表达,提示其恶性不断升高;ＴＧＦβＲＩＩ 表达不断减弱,促进胶质瘤发生和发展。     

bFGF基因表达与人脑胶质瘤间质血管增生

探讨ｂＦＧＦ基因表达与人脑胶质瘤间质血管增生的关系。方法采用原位杂交法检测５７例胶质瘤组织的ｂＦＧＦｍＲＮＡ表达水平并定量分析所含毛细血管。结果：各级胶质瘤ｂＦＧＦｍＲＮＡ表达的阳性性,随着胶质瘤及别的升达水平也升高。     

膀胱移行细胞癌中bFGF的表达及其与微血管定量的关系

目的：为了探讨膀胱移行细胞癌中碱性成纤维细胞生长因子（ｂＦＧＦ）的表达及其与血管形成定量的关系。方法：应用免疫组织化学方法,对６２例原发性膀胱移行细胞癌组织中ｂＦＧＦ进行检测,并对浸润性膀胱癌组织中微血管进行定量研究。结果：膀胱癌中ｂＦＧＦ阳性表达率为４６．８％,与病理分级、临床分期、复发及预后有关（Ｐ〈０．０５）,与浸润性癌组织中微血管定量呈正相关（Ｐ〈０．０５）,结论：ｂＦＧＦ表达对膀胱癌生物学行为有重要影响,ｂＦＧＦ是膀胱癌血管形成过程中一个主要血管生成因子,ｂＦＧＦ表达和血管形成的定量有可能成为预测膀胱癌复发、转移和预后的重要指标。     

bFGF在肾癌中表达的意义

应用免疫组化方法检测４１例肾癌和１１例正常肾组织中ｂＦＧＦ的表达。结果显示：肾癌组织中ｂＦＧＦ的阳性率为７５．６％（３１／４１），正常肾组织为５４．５％（６／１１），两者相比无显著性差异，两者的强阳性率分别为６１．０％、２７．３％，有显著性差异。ｂＦＧＦ表达的强度与肾癌病理分级、分期密切相关。提示ｂＦＧＦ参与肾癌的发生发展过程，可作为肾癌生物学行为的一项评估指标。     

创伤愈合中肥大细胞的形态变化及定量研究

创伤愈合过程中，参与或涉及的种类很多，其中肥大细胞、成纤维细胞等对创伤愈合起着极为重要的作用。本实验以雄性北京小型猪为创伤模型，观察了碱性成纤维细胞生长因子（ｂＦＧＦ）治疗后肥大细胞（Ｍａｓｔｃｅｌ，ＭＣ）在创伤愈合中的动态变化，并结合图像分析进行了定量研究。结果发现：创伤后ＭＣ数量和ＭＣ内颗粒数量发生了改变，ｂＦＧＦ也参与了创伤修复过程。本研究的结果提示ｂＦＧＦ和ＭＣ可能参与了这一过程，但对于这两者的关系和在其中的作用均需要进一步研究。     

表皮生长因子受体反义RNA抑制胶质瘤细胞生长增殖的体外试验

目的：研究表皮生长因子受体基因对于胶质瘤发生发展的作用，探索以ＥＧＦＲ反义ＲＮＡ进行反义基因治疗胶质瘤的可行性。方法：将含ＥＧＦＲ反义ＲＮＡ的质粒通过脂质体介导转入Ｃ６恶性胶质瘤细胞，通过Ｓｏｕｔｈｅｒｎ印迹杂交鉴定外源基因整合情况，ＭＴＴ法测定细胞存活率，Ｎｏｒｔｈｅｒｎ印迹杂交、细胞原位ｍＲＮＡ杂交及ＥＧＦＲ免疫组化染色检测ＥＧＦＲｍＲＮＡ及蛋白表达，核仁组成区相关嗜银蛋白染色（ＡｇＮＯＲ计数）检测细胞增殖活性。结果：Ｓｏｕｔｈｅｒｎ印迹杂交表明外源性反义ＥＧＦＲ基因（互补于ＥＧＦＲ基因全长及３端部分序列）分别在Ｃ６细胞中整合（分别命名为Ｃ－ｐＲ１和Ｃ－ｐＲ３），通过Ｎｏｒｔｈｅｒｎ印迹杂交、原位杂交及细胞免疫组织化学检测均显示转染ＥＧＦＲ反义ＲＮＡ的Ｃ６细胞比转染前ＥＧＦＲｍＲＮＡ水平及ＥＧＦＲ蛋白水平有不同程度的降低。表达高水平反义ＲＮＡ的克隆在培养状态下细胞增殖活性较对照组有明显下降。结论：ＥＧＦＲ在恶性胶质瘤细胞的发生发展过程中起十分关键的作用，ＥＧＦＲ有可能成为基因治疗恶性胶质瘤的优选靶的之一。     

乳腺癌c—erbB—2,表皮生长因子受体基因蛋白表达与预后的关系

以免疫组化ＡＢＣ法检测乳腺癌ｃ－ｅｒｂＢ－２、表皮生长因子受体基因蛋白表达，研究其与预后的关系，结果：ｃ－ｅｒｂＢ－２、ＥＧＦＲ阳性表达率各为３８．５％（２５／６５）和４３．１％（２８／６５）。除ＥＧＦＲ表达与ＥＲ、ＰＲ负相关外，ｃ－ｅｒｂＢ－２，ＥＧＦＲ表达与临床预后因素无相关性，二者之间亦无相关性。ｃ－ｅｒｂＢ－２或ＥＧＦＲ阳性组术后生存显著差于阴性组（Ｐ〈０．０１）；多因素分析二者是显著影响     

ＧＦＩ１Ｂ在急性白血病细胞中的表达及临床意义

目的　观察急性白血病（ＡＬ）细胞的独立生长因子１Ｂ（ＧＦＩ１Ｂ）的表达水平及临床意义。方法　采用实时荧光定量ＰＣＲ（ＲＱ-ＰＣＲ）检测急性髓系白血病（ＡＭＬ）患者４８例、急性淋巴细胞白血病（ＡＬＬ）患者７例和正常对照１５例中ＧＦＩ１Ｂ的表达。结果　（１）在ＡＭＬ组,初治ＡＭＬ患者ＧＦＩ１Ｂ平均表达水平较正常对照组明显升高,差异有显著性（Ｐ＜０.０５）,经治疗获得完全缓解的患者ＧＦＩ１Ｂ平均表达水平较治疗前明显下降（Ｐ＜０.０５）,病情复发的患者ＧＦＩ１Ｂ平均表达水平再度升高;（２）在初治ＡＭＬ的Ｍ１～Ｍ７各亚型间ＧＦＩ１Ｂ平均表达水平有差别,Ｍ１～Ｍ５亚型ＧＦＩ１Ｂ表达水平略高于正常对照组,但差异均无显著性（Ｐ＞０.０５）,在Ｍ６和Ｍ７亚型ＧＦＩ１Ｂ平均表达水平均明显高于正常对照组,差异均有显著性（Ｐ＜０.０５）;（３）在ＡＬＬ组,初治ＡＬＬ患者、ＡＬＬ复发患者以及完全缓解的患者ＧＦＩ１Ｂ平均表达水平与正常对照组比较均无明显改变,差异均无显著性（Ｐ＞０.０５）。结论　ＧＦＩ１Ｂ在ＡＬ患者中有不同的表达,并提示其与ＡＭＬ,尤其是Ｍ６和Ｍ７亚型的发病及病情变化密切相关。     

胶质瘤中FGFR1OP和p57/Kip2蛋白的临床意义

目的:探讨FGFR1OP和p57/Kip2在胶质瘤发生、发展中的作用和临床意义.方法:对中国医科大学附属盛京医院和北京通州区潞河医院的54例胶质瘤手术切除标本,采用SP法进行免疫组织化学染色,检测FGFR1OP和p57/Kip2的表达情况,分析其与胶质瘤临床病理各参数的相关性,并评价其临床意义.结果:FGFR1OP和p57/Kip2在胶质瘤中的阳性表达率分别为66.7%和44.4%,且随着胶质瘤的恶性程度不同表现出显著性差异(P<0.05),FGFR1OP在高度恶性胶质瘤(Ⅲ～Ⅳ级)中的OD值为0.131±0.010,高于在低度恶性胶质瘤(Ⅰ～Ⅱ级)中的OD值(0.118±0.010),两者有显著统计学差异(P=0.000).p57/Kip2在高度恶性胶质瘤(Ⅲ～Ⅳ级)中的OD值(0.156±0.008),低于在低度恶性胶质瘤(Ⅰ～Ⅱ级)中的OD值(0.165±0.006),两者有显著统计学差异(P=0.014).Pearson相关系数检验分析提示FGFR10P和p57/Kip2之间呈负直线相关(r=-0.732,P<0.01).结论:FGFR1OP的高表达与p57/Kip2的低表达可能参与胶质瘤的生长、分化和进展,提示预后不良.     

脑膜瘤组织中bFGF及FGFR-1蛋白表达的研究

目的　探讨碱性成纤维生长因子 (basicfibroblastgrowthfactor ,bFGF)及成纤维生长因子受体 1(fibroblastgrowthfactorreceptor 1,FGFR 1)在脑膜瘤中的表达及其与脑膜瘤组织病理学和复发的关系。方法　用免疫组化技术检测bFGF、FGFR 1在不同类型的脑膜瘤组织中的蛋白表达 ,用组织病理学判断脑膜瘤的良恶性。结果　脑膜瘤细胞有不同程度的bFGF及FGFR 1表达 ,其表达阳性率与肿瘤的良恶性和复发有关。结论　bFGF和FGFR具有促进脑膜瘤细胞的增殖和生长的作用。脑膜瘤表达bFGF、FGFR的阳性率可作为鉴别肿瘤良恶性的有用指标 ,并对脑膜瘤的预后和术后复发起提示作用。     

Th2类细胞因子优势表达在人脑胶质瘤发生及发展中的作用

目的 探讨Th1／Th2类细胞因子基因在人脑胶质瘤中的分泌及表达情况,以及它们在脑胶质瘤发生及发展中的作用。方法 以IL—2、IFNγ代表Th1类细胞因子,IL—4、IL—6及IL—10代表Th2类细胞因子,采用ELISA法检测脑胶质瘤细胞株培养上清中细胞因子的活性,采用逆转录聚合酶链反应方法(RT—PCR)检测Th1／Th2类细胞因子基因在脑胶质瘤细胞、肿瘤浸润淋巴细胞及细胞株中的表达情况。结果 人脑胶质瘤细胞、肿瘤浸润淋巴细胞及脑胶质瘤细胞株均呈现明显的Th2类细胞因子基因优势表达,而正常人脑组织中则无这种表达趋势。结论 Th2类细胞因子在人脑胶质瘤中的优势表达可能是导致脑胶质瘤患者细胞免疫功能低下的原因之一,有可能在脑胶质瘤的发生及发展中起一定作用。     

Immunohistochemical expression of basic fibroblast growth factor and fibroblast growth factor receptors 1 and 2 in the pathogenesis of lung cancer

PURPOSE: To identify the patterns of protein expression of basic fibroblast growth factor (bFGF) and FGF receptors 1 and 2 in non-small cell lung carcinoma (NSCLC) and their role in the early pathogenesis of squamous cell carcinoma (SCC) of the lung. EXPERIMENTAL DESIGN: Archived tissue from NSCLC (adenocarcinoma and SCC; n = 321) and adjacent bronchial epithelial specimens (n = 426) were analyzed for the immunohistochemical expression of bFGF, FGFR1, and FGFR2, and the findings were correlated with clinicopathologic features of the patients. RESULTS: High expression of bFGF, FGFR1, and FGFR2 was shown in most NSCLC tumors. The pattern of expression for all markers varied according to tumor histologic type and cellular localization. Cytoplasmic expression scores were significantly higher in tumors than in normal epithelia. Nuclear bFGF (P = 0.03) and FGFR1 (P = 0.02) levels were significantly higher in women than in men. Although cytoplasmic FGFR1 expression was significantly higher (P = 0.002) in ever smokers than in never smokers, nuclear FGFR1 (P = 0.0001) and FGFR2 (P = 0.003) expression was significantly higher in never smokers. Different prognostic patterns for the expression of these markers were detected for both NSCLC histologic types. Dysplastic changes showed significantly higher expression of all markers compared with squamous metaplasia. CONCLUSIONS: bFGF, FGFR1, and FGFR2 are frequently overexpressed in SCC and adenocarcinoma of the lung. bFGF signaling pathway activation may be an early phenomenon in the pathogenesis of SCC and thus an attractive novel target for lung cancer chemopreventive and therapeutic strategies.     

连接蛋白基因Cx43抑制胶质瘤细胞增殖及其机理的初步探讨

目的 探讨连接蛋白基因Cx43对胶质瘤细胞增殖的抑制及其可能的机理。方法 将含Cx43cDNA的质粒以脂质体介导转染Cx43表达缺失的人和鼠的恶性胶质瘤细胞,通过Northem杂交、原位杂交及免疫组化染色检测Cx43mRNA及蛋白表达;MTT法测定细胞增殖率;核仁组成区嗜银蛋白染色检测细胞增殖活性;TUNEL法检测细胞凋亡;划痕标记荧光染料示踪技术检测细胞间隙连接通讯(GJIC);Western杂交及免疫组化染色检测bFGF、PDGF、EGFR、IGF-I和IGFBP3的表达。结果 转染Cx43基因的胶质瘤细胞增殖下降,GJIC恢复,同时伴有bFGF、PDGF、IGF-I和IGFBP3表达下降,而EGFR表达和细胞凋亡则无改变。结论 Cx43基因可能通过恢复GJIC功能及抑制某些重要生长因子的自分泌,实现对胶质瘤细胞增殖的抑制。     

Inhibition of autocrine fibroblast growth factor signaling by the adenovirus-mediated expression of an antisense transgene or a dominant negative receptor in human glioma cells in vitro

Autocrine fibroblast growth factor (FGF) signaling was genetically manipulated in human gliomas by an adenovirus-mediated strategy. Antisense inhibition of endogenous basic FGF (bFGF) expression showed inhibition of the proliferation of human glioma cells (U251MG, NMC-G1) without promoting apoptosis. The reduction in proliferation in response to antisense inhibition was reversed with an additional supplement of exogenous bFGF. On the other hand, the overexpression of a kinase defective mutant of the type I FGF receptor (dominant negative FGF receptor) resulted in not only growth inhibition but also marked apoptosis in U251MG cells. In the cells expressing a dominant negative FGF receptor, the reduction in cell number was not reversed by the exogenous bFGF supplement. Our data further implicated the significance of an autocrine FGF signaling loop in human gliomas. Cell survival activity may largely depend on the receptor-mediated pathway, which should be considered in the development of the molecular based therapeutics.     

Adenovirus-mediated transfer of siRNA against basic fibroblast growth factor mRNA enhances the sensitivity of glioblastoma cells to chemotherapy

Basic fibroblast growth factor (bFGF) is an important growth factor for glioma cell proliferation and invasion. BFGF is overexpressed in malignant gliomas and its level is associated with malignant grades and clinical outcome of patients with gliomas. Small interfering RNAs (siRNA) are synthetic forms of microRNA made of short double stranded RNA, and they effectively catalyze the degradation of their target mRNA. The use of siRNA has become a key method in the suppression of gene expression and the development of therapeutic agents. In this study, we used an adenovirus(Ad)-mediated transfer of siRNA against bFGF mRNA (Ad-bFGF-siRNA) to study the effect of down-regulating bFGF expression on the sensitivity of glioma cells to chemotherapeutics. An optimal siRNA sequence specific for bFGF mRNA was cloned into an adenoviral vector and transfected into three glioma cell lines: U251, A172, and LN229. Methyl thiazolyl tetrazolium (MTT) assays were used to examine changes in cell proliferation, and changes in bFGF mRNA and protein levels in U251 cells were detected using quantitative RT-PCR and Western blot, respectively. Apoptosis of U251 cells was detected using Hoechst staining and flow cytometry, with expression of apoptosis-related proteins evaluated by Western blot. Following the transfection of a bFGF-specific siRNA, mRNA and protein levels of bFGF decreased significantly. Lower rates of proliferation and increased levels of apoptosis also were associated with the Ad-bFGF-siRNA-transfected group compared to control group. Decreased expression of Bcl-2, Bcl-xL, Jak-1, and STAT-3 and increased expression of Bax also were detected in the Ad-bFGF-siRNA-transfected group. For cells treated with both Ad-bFGF-siRNA and chemotherapeutics, a significant reduction in cell survival was observed compared to treatment with Ad-bFGF-siRNA or chemotherapeutics alone. Overall, we found that targeting bFGF mRNA with a siRNA resulted in lower rates of proliferation, increased apoptosis, and enhanced sensitivity of glioma cells to chemotherapy drugs. This suggests that specific targeting of bFGF mRNA may provide a novel approach for the treatment of glioblastoma multiforme (GBM).     

碱性成纤维细胞生长因子在不同级别人脑胶质瘤中的表达

目的:研究碱性成纤维细胞生长因子(bFGF)与人脑胶质瘤恶性程度之间的关系。方法:取手术切除人脑胶质瘤标本45例,手术内减压正常脑组织5例,免疫组织化学染色观察bFGF在正常脑组织及不同级别脑胶质瘤组织中的表达水平。结果:正常脑组织无bFGF表达,随胶质瘤级别增加bFGF表达率明显增高,正常脑组织、Ⅰ、Ⅱ、Ⅲ、Ⅳ级胶质瘤两两之间差异有显著性(P<0.05)。结论:bFGF在正常脑组织中不表达,其表达与人脑胶质瘤的恶性程度正相关,可成为判断胶质瘤恶性程度和预后的一项指标,并为胶质瘤的治疗提供新的途径。