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中国误诊学杂志 2011年第31期11卷 7575-7576页

作者：沈利军孙冬霞李文静杜丽敏河北省邯郸市眼科医院病理科 056001 邯郸市妇幼保健院病理科邯郸市眼科医院检验科

目的检测子宫内膜癌组织中HER-2、PTEN蛋白表达。方法采用免疫组化法检测60例子宫内膜癌、32例正常子宫内膜中HER-2、PTEN的表达。结果子宫内膜癌中HER-2的表达率高于正常子宫内膜(P<0.05),且与肿瘤分级、分期以及浸润深度有关(P<... 详细信息

目的检测子宫内膜癌组织中HER-2、PTEN蛋白表达。方法采用免疫组化法检测60例子宫内膜癌、32例正常子宫内膜中HER-2、PTEN的表达。结果子宫内膜癌中HER-2的表达率高于正常子宫内膜(P<0.05),且与肿瘤分级、分期以及浸润深度有关(P<0.05);内膜癌中PTEN表达缺失率高于正常宫内膜(P<0.05)。PTEN缺失率与肿瘤分级以及类型有关(P<0.05)。HER-2与PTEN的表达呈负相关(r=-0.698,P<0.05)。结论 HER-2与PTEN的联合检测有助于子宫内膜癌的早期诊断及预后评估。

关键词：子宫内膜肿瘤/代谢表皮生长因子/代谢 PTEN磷酸水解酶/代谢

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环氧化酶-2和表皮生长因子受体在星形细胞瘤中表达的意义

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中国医师杂志 2010年第2期12卷 185-187页

作者：张洪涛田苗修春明刘杰崔广强王云波潘树茂青岛大学医学院附属烟台毓璜顶医院神经外科山东省烟台264000

目的探讨环氧化酶-2（COX-2）、表皮生长因子受体（EGFR）在星形细胞瘤中的表达及其相关性。方法用免疫组织化学s—p法检测68例不同病理级别星形细胞瘤组织与5例正常脑组织中COX-2、EGFR及PCNA蛋白的表达，并计算增殖指数（PI），分析CO... 详细信息

目的探讨环氧化酶-2（COX-2）、表皮生长因子受体（EGFR）在星形细胞瘤中的表达及其相关性。方法用免疫组织化学s—p法检测68例不同病理级别星形细胞瘤组织与5例正常脑组织中COX-2、EGFR及PCNA蛋白的表达，并计算增殖指数（PI），分析COX-2表达与EGFR及PI之间的相关性。结果正常脑组织中COX-2、EGFR表达均为阴性，高度恶性组的COX-2阳性表达率明显高于低度恶性组（73．53％vs 44．18％，P〈0．01），高度恶性组PI（％）明显高于低度恶性组和对照组（46．11±10．68VS23．0g4-6．25，4．52±0．95，P〈0．01）。在低度恶性组与高度恶性组中，COX-2表达阳性的肿瘤其PI明显高于阴性者（P〈0．01）。高度恶性组的EGFR阳性表达率明显高于低度恶性组（67．65％vs38．24％，P〈0．01）。COX-2蛋白在EGFR阳性组的表达明显高于阴性组。结论COX-2、EGFR的表达与星形细胞瘤的病理分级有关，COX-2可促进肿瘤细胞增殖，并与EGFR的表达密切相关，EGFR信号通路可能参与COX-2表达的调控。

关键词：环氧化酶2/代谢受体表皮生长因子/代谢星形细胞瘤/代谢 ’

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脑胶质瘤60例CXCR_4与EGFR的表达及意义

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陕西医学杂志 2008年第9期37卷 1226-1228页

作者：王东林刘毅高宝莲南通大学医学院病理学教研室南通226001

目的:探讨CXCR4和EGFR在脑胶质瘤中的表达状况及其与肿瘤发生、发展和侵袭性生长的关系。方法:采用免疫组织化学SP法,检测60例脑胶质瘤、5例正常脑组织中CXCR4和EGFR的表达。用统计学方法分析其表达及与胶质瘤病理分级的关系。结果:60... 详细信息

目的:探讨CXCR4和EGFR在脑胶质瘤中的表达状况及其与肿瘤发生、发展和侵袭性生长的关系。方法:采用免疫组织化学SP法,检测60例脑胶质瘤、5例正常脑组织中CXCR4和EGFR的表达。用统计学方法分析其表达及与胶质瘤病理分级的关系。结果:60例胶质瘤患者中分别有39例表达CXCR4和EGFR。CXCR4在Ⅱ、Ⅲ及Ⅳ级胶质瘤中阳性表达率分别为38.10%、73.91%、87.50%;EGFR在Ⅱ、Ⅲ及Ⅳ级胶质瘤中阳性表达率分别为52.38%、60.87%、87.50%。正常脑组织均不表达CXCR4、EGFR。CXCR4、EGFR在不同级别胶质瘤间的表达有显著性差异(P<0.05),两者随着胶质瘤病理分级增高而表达增强(rs=0.426,P<0.05)。结论:CXCR4与EGFR表达呈正相关,均与胶质瘤的病理分级有关。

关键词：脑肿瘤/病理学受体趋化因子/代谢表皮生长因子/代谢免疫组织化学

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Requirement of SHP2 binding to Grb2-associated binder-1 for mitogen-activated protein kinase activation in response to lysophosphatidic acid and epidermal growth factor

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第18期275卷 13842-13848页

作者： Cunnick, JM Dorsey, JF Munoz-Antonia, T Mei, L Wu, J H Lee Moffitt Canc Ctr & Res Inst Mol Oncol Program MRC 3057E Tampa FL 33612 USA Univ S Florida Dept Med Microbiol & Immunol Tampa FL 33612 USA Univ S Florida Dept Biochem & Mol Biol Tampa FL 33612 USA Univ Alabama Birmingham Dept Neurobiol Birmingham AL 35294 USA

Grb2-associated binder-1 (Gab1) is a multisite docking protein containing a pleckstrin homology (PH) domain, multiple potential tyrosine phosphorylation sites, and several proline-rich sequences. Gab1 becomes tyrosine-phosphorylated in cells stimulated with growth factors, cytokines, and ligands for G protein-coupled receptors, A major Gab1-binding protein detected in cells treated with extracellular stimuli is the tyrosine phosphatase, SHP2. Although the role of SHP2-Gab1 interaction in cell signaling has not yet been characterized, SHP2 is known to mediate mitogen-activated protein (MAP) kinase activation induced by the epidermal growth factor (EGF), However, the mechanism by which the SHP2 phosphatase exerts a positive signaling role remains obscure. In this study, we prepared Gab1 mutants lacking the SHP2 binding site (Gab1Y627F), the phosphatidylinositol 3-kinase (PI3K) binding sites (Gab1 Delta PI3K), and the PH domain (Gab1 Delta PH). Expression of Gab1Y627F blocked the extracellular signal-regulated kinase-2 (ERK2) activation by lysophosphatidic acid (LPA) and EGF, Conversely, expression of the wild-type Gab1 in HEK293 cells augmented the LPA receptor Edg2-mediated ERK2 activation. Whereas the PH domain was required for Gab1 mediation of ERR2 activation by LPA, it was not essential for EGF-induced ERK2 activation. Expression of Gab1 Delta PI3K had no apparent effect on ERK2 activation by LPA and EGF in the cells that we have examined. These results establish a role for Gab1 in the LPA-induced MAP kinase pathway and clearly demonstrate that Gab1-SHP2 interaction is essential for ERK2 activation by LPA and EGF. These findings also suggest that the positive role of SHP2 in the MAP kinase pathway depends on its interaction with Gab1.

关键词： COS细胞酶激活表皮生长因子/代谢表皮生长因子/药理学细胞内信号肽和蛋白质类溶血磷脂素类/代谢溶血磷脂素类/药理学丝裂原激活蛋白激酶类/代谢磷蛋白类/代谢蛋白质结合蛋白质酪氨酸磷酸酶非受体11型蛋白酪氨酸磷酸酶非受体6型蛋白质酪氨酸磷酸酶类/代谢信号传导/药物作用动物

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Tenascin-R is a functional modulator of sodium channel β subunits

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JOURNAL OF BIOLOGICAL CHEMISTRY 1999年第37期274卷 26511-26517页

作者： Xiao, ZC Ragsdale, DS Malhotra, JD Mattei, LN Braun, PE Schachner, M Isom, LL Univ Michigan Dept Pharmacol Ann Arbor MI 48109 USA McGill Univ Montreal Neurol Inst Montreal PQ H3A 2B4 Canada McGill Univ Dept Biochem Montreal PQ H3G 1Y6 Canada Univ Hamburg Zentrum Mol Neurobiol D-20246 Hamburg Germany

Voltage-gated sodium channels isolated from mammalian brain are composed of a, beta 1 and beta 2 subunits, The cu subunit forms the ion conducting pore of the channel, whereas the beta 1 and beta 2 subunits modulate channel function, as well as channel plasma membrane expression levels, beta 1 and beta 2 each contain a single, extracellular Ig-like domain with structural similarity to the neural cell adhesion molecule (CAM), myelin Po. beta 2 contains strong amino acid homology to the third Ig domain and to the juxtamembrane region of F3/contactin. Many CAMs of the Ig superfamily have been shown to interact with extracellular matrix molecules. We hypothesized that beta 2 may interact with tenascin-alpha (TN-R), an extracellular matrix molecule that is secreted by oligodendrocytes during myelination and that binds F3-contactin. We show here that cells expressing sodium channel beta 1 or beta 2 subunits are functionally modulated by TN-R, Transfected cells stably expressing beta 1 or beta 2 subunits initially recognized and then were repelled from TN-R substrates, The cysteine-rich amino-terminal domain of TN-R expressed as a recombinant peptide, termed EGF-L, appears to be responsible for the repellent effect on beta subunit-expressing cells. The epidermal growth factor-like repeats and fibronectin-like repeats 6-8 are most effective in the initial adhesion of id subunit-expressing cells. Application of EGF-L to alpha IIA beta 1 beta 2 channels expressed in Xenopus oocytes potentiated expressed sodium currents without significantly altering current time course or the voltage dependence of current activation or inactivation. Thus, sodium channel beta subunits appear to function as CAMs, and TN-R may be an important regulator of sodium channel localization and function in neurons.

关键词：氨基酸序列细胞黏附细胞系仓鼠亚科仓鼠属表皮生长因子/代谢分子序列数据重组蛋白质类/化学重组蛋白质类/遗传学重组蛋白质类/代谢钠通道/化学钠通道/代谢腱糖蛋白/化学腱糖蛋白/遗传学腱糖蛋白/代谢转染爪蟾属动物小鼠

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Aging fibroblasts present reduced epidermal growth factor (EGF) responsiveness due to preferential loss of EGF receptors

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第25期275卷 19343-19351页

作者： Shiraha, H Gupta, K Drabik, K Wells, A Univ Pittsburgh Dept Pathol Pittsburgh PA 15261 USA Univ Alabama Birmingham Dept Pathol Birmingham AL 35294 USA

Wound healing is compromised in aging adults in part due to decreased responsiveness of fibroblasts to extracellular signals. However, the cellular mechanisms underlying this phenomenon are not known. Aged dermal fibroblasts with reduced remaining replicative capacities demonstrated decreased epidermal growth factor (EGF)-induced cell migrative and cell proliferative capacities, as reported previously. Thus, as cells approach senescence, programmed in vivo or in vitro, EGF responsiveness is preferentially lost. To define the rate-limiting signaling event, we found that the activity of two different EGF receptor (EGFR)-signaling pathways to cell migration (phospholipase-C gamma) and/or mitogenesis (extracellular signal/regulated-mitogen-activated kinases) were decreased in near senescent cells despite unchanged levels of effector molecules. Aged cells presented decreased levels of EGFR, although insulin receptor and transferrin receptor levels were relatively unchanged. EGFR, mRNA levels and production of new transcripts decreased during aging, suggesting that this preferential loss of EGFR was due to diminished production, which more than counteracts the reduced ligand-induced receptor loss. Since these data suggested that the decrement in EGF was rate-limiting, higher levels of EGFR were established in near senescent cells by electroporation of EGFR cDNA. These cells presented higher levels of EGFR and recovered their EGF-induced migration and proliferation responsiveness. Thus, the defect in EGF responsiveness of aged dermal fibroblasts is secondary to reduced EGFR message transcription. Our experimental model suggests that EGFR gene delivery might be an effective future therapy for compromised wound healing.

关键词：细胞衰老细胞分裂细胞运动环AMP/代谢 DNA 互补表皮生长因子/代谢成纤维细胞/细胞学成纤维细胞/代谢启动区(遗传学) 受体表皮生长因子/遗传学受体表皮生长因子/代谢信号传导人类

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Selective perturbation of early endosome and/or trans-Golgi network pH but not lysosome pH by dose-dependent expression of influenza M2 protein

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JOURNAL OF BIOLOGICAL CHEMISTRY 1999年第14期274卷 9854-9860页

作者： Henkel, JR Popovich, JL Gibson, GA Watkins, SC Weisz, OA Univ Pittsburgh Renal Electrolyte Div Lab Epithelial Cell Biol Pittsburgh PA 15213 USA Univ Pittsburgh Dept Cell Biol & Physiol Pittsburgh PA 15213 USA

Many sorting stations along the biosynthetic and endocytic pathways are acidified, suggesting a role for pH regulation in protein traffic, However, the function of acidification in individual compartments has been difficult to examine because global pH perturbants affect all acidified organelles in the cell and also have numerous side effects, To circumvent this problem, we have developed a method to selectively perturb the pH of a subset of acidified compartments, We infected HeLa cells with a recombinant adenovirus encoding influenza virus M2 protein (an acid-activated ion channel that dissipates proton gradients across membranes) and measured the effects on various steps in protein transport, Al low multiplicity of infection (m.o.i.), delivery of influenza hemagglutinin from the trans-Golgi network to the cell surface was blocked, but there was almost no effect on the rate of recycling of internalized transferrin, At higher m.o.i., transferrin recycling was inhibited, suggesting increased accumulation of M2 in endosomes, Interestingly, even at the higher m.o.i., M2 expression had no effect on lysosome morphology or on EGF degradation, suggesting that lysosomal pH was not compromised by M2 expression. However, delivery of newly synthesized cathepsin D to lysosomes was slowed in cells expressing active M2, suggesting that acidification of the TGN and endosomes is important for efficient delivery of lysosomal hydrolases, Fluorescence labeling using a pH-sensitive dye confirmed the reversible effect of M2 on the pH of a subset of acidified compartments in the cell, The ability to dissect the role of acidification in individual steps of a complex pathway should be useful for numerous other studies on protein processing and transport.

关键词：腺病毒科生物转运组织蛋白酶D/代谢细胞区室化细胞极性剂量效应关系药物内颗粒/代谢表皮生长因子/代谢遗传载体高尔基体/代谢 HeLa细胞氢离子浓度流感病毒A型/遗传学流感病毒A型/代谢离子通道/遗传学离子通道/代谢溶酶体/代谢病毒基质蛋白质类/生物合成人类

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The protein-tyrosine phosphatase SHP-1 binds to and dephosphorylates p120 catenin

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第34期275卷 26376-26384页

作者： Keilhack, H Hellman, U van Hengel, J van Roy, F Godovac-Zimmermann, J Böhmer, FD Univ Jena Klinikum Res Unit Mol Cell Biol D-07747 Jena Germany Ludwig Inst Canc Res Uppsala Branch SE-75124 Uppsala Sweden State Univ Ghent VIB Dept Mol Biol Mol Cell Biol Unit B-9000 Ghent Belgium Inst Mol Biotechnol D-07745 Jena Germany

A prominent tyrosine-phosphorylated protein of similar to 100 kDa (designated pp100) in epidermal growth factor (EGF)-stimulated A431 cells was found to be a main interaction partner of the protein-tyrosine phosphatase SHP-1 in pull-down experiments with a glutathione S-transferase-SHP-1 fusion protein. Binding was largely mediated by the N-terminal SH2 domain of SHP-1 and apparently direct and independent from the previously described association of SHP-1 with the activated EGF receptor. pp100 was partially purified and identified by mass spectrometric analysis of tryptic fragments, partial amino acid sequencing, and use of authentic antibodies as the 3A isoform of the Armadillo repeat protein superfamily member p120 catenin (p120(ctn)). Different p120(ctn) isoforms expressed in human embryonal kidney 293 cells, exhibited differential binding to SHP-1 that correlated partly with the extent of EGF-dependent p120(ctn) tyrosine phosphorylation. Despite strong phosphorylation, p120(ctn) isoforms 3B and 3AB bound, however, less readily to SHP-1. SHP-1 associated transiently with p120(ctn) in EGF-stimulated A431 cells stably transfected with a tetracycline-responsive SHP-1 expression construct, and p120(ctn) exhibited elevated phosphorylation upon a tetracycline-mediated decrease in the SHP-1 level. Functions of p120(ctn), which are regulated by tyrosine phosphorylation, may be modulated by the described SHP-1-p120(ctn) interaction.

关键词：细胞黏附分子/代谢细胞系电泳聚丙烯酰氨凝胶表皮生长因子/代谢细胞内信号肽和蛋白质类分子量磷蛋白类/代谢磷酰化蛋白质结合蛋白质酪氨酸磷酸酶非受体11型蛋白酪氨酸磷酸酶非受体6型蛋白质酪氨酸磷酸酶类/代谢含SH2域蛋白质酪氨酸磷酸酶类光谱法质量基质辅助激光解吸电离 src同源域人类

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Polyubiquitination of the epidermal growth factor receptor occurs at the plasma membrane upon ligand-induced activation

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第18期275卷 13940-13947页

作者： Stang, E Johannessen, LE Knardal, SL Madshus, IH Univ Oslo Natl Hosp Inst Pathol N-0027 Oslo Norway

We have previously shown that, although overexpression of mutant dynamin inhibits clathrin-dependent endocytosis and disrupts high affinity binding of epidermal growth factor (EGF) to the EGF receptor (EGFR), it does not inhibit ligand-induced translocation of the EGFR into clathrin-coated pits. In the present study, we demonstrate that, upon ligand binding and incubation at 37 degrees C, the EGFR was polyubiquitinated regardless of overexpression of mutant dynamin, In cells not overexpressing mutant dynamin, the EGFR was rapidly internalized and deubiquitinated, In cells being endocytosis-deficient, due to overexpression of mutant dynamin, however, the EGFR was upon prolonged chase first found in deeply invaginated coated pits, and then eventually moved out of the coated pits and back onto the smooth plasma membrane, Polyubiquitination occurred equally efficiently in cells with or without intact clathrin-dependent endocytosis, while the kinetics of ubiquitination and deubiquitination was somewhat different. We further found that the EGF-induced ubiquitination of Eps15 occurred both in the absence and presence of endocytosis with the same kinetics as polyubiquitination of the EGFR, but that the EGF-induced monoubiquitination of Eps15 was somewhat reduced upon overexpression of mutant dynamin, Our data show that EGF-induced polyubiquitination of the EGFR occurs at the plasma membrane.

关键词：细胞膜/代谢动力蛋白质类表皮生长因子/代谢 GTP磷酸水解酶类/遗传学 GTP磷酸水解酶类/代谢 HeLa细胞配体突变信号传导泛素类人类

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The evolutionarily conserved n-terminal region of Cbl is sufficient to enhance down-regulation of the epidermal growth factor receptor

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第1期275卷 367-377页

作者： Lill, NL Douillard, P Awwad, RA Ota, S Lupher, ML Miyake, S Meissner-Lula, N Hsu, VW Band, H Harvard Univ Brigham & Womens Hosp Sch MedLymphocyte Biol Sect Dept MedDiv Rheumatol Immunol & Allergy Boston MA 02115 USA

The mammalian proto-oncoprotein Cbl and its homologues in Caenorhabditis elegans and Drosophila are evolutionarily conserved negative regulators of the epidermal growth factor receptor (EGF-R), Overexpression of wild-type Cbl enhances down-regulation of activated EGF-R from the cell surface. We report that the Cbl tyrosine kinase-binding (TKB) domain is essential for this activity. Whereas wild-type Cbl enhanced ligand-dependent EGF-R ubiquitination, down-regulation from the cell surface, accumulation in intracellular vesicles, and degradation, a Cbl TKB domain-inactivated mutant (G306E) did not. Furthermore, the transforming truncation mutant Cbl-N (residues 1-357), comprising only the Cbl TKB domain, functioned as a dominant negative protein. It colocalized with EGF-R in intracellular vesicular structures, yet it suppressed down-regulation of EGF-R from the surface of cells expressing endogenous wild-type Cbl. Therefore, Chi-mediated down-regulation of EGF-R requires the integrity of both the N-terminal TKB domain and additional C-terminal sequences. A Cbl truncation mutant comprising amino acids 1-440 functioned like wild-type Cbl in down-regulation assays. This mutant includes the evolutionarily conserved TKB and RING finger domains but lacks the less conserved C-terminal sequences, We conclude that the evolutionarily conserved N terminus of Cbl is sufficient to effect enhancement of EGF-R ubiquitination and down-regulation from the cell surface.

关键词：氨基酸序列生物转运保守序列减量调节胞吞作用表皮生长因子/代谢绿色荧光蛋白质类血凝素类/遗传学血凝素类/代谢细胞内膜/代谢发光蛋白质类/遗传学发光蛋白质类/代谢突变肽碎片/遗传学肽碎片/代谢磷酰化蛋白质结合蛋白质结构三级蛋白酪氨酸激酶类/代谢原癌基因蛋白质类/遗传学原癌基因蛋白质类/代谢原癌基因蛋白质c-cbl 重组融合蛋白质类/代谢酪氨酸/代谢泛素蛋白连接酶类泛素类/代谢

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