粒细胞集落刺激因子及其受体

粒细胞集落刺激因子（Ｇ－ＣＳＦ）是一种多肽链的细胞生长因子,可特异地调节粒系细胞的增殖与分化,并能增强成熟粒细胞的功能,对机体应激防御系统有重要意义。近年来发现Ｇ－ＣＳＦ与白血病细胞的凋亡有一定关系。Ｇ－ＣＳＦ功能的发挥有赖于与效应细胞表面的特异性受体的结合。随着基因克隆技术的发展,Ｇ－ＣＳＦ重组产品已广泛用于临床,为血液病及其他疾病的治疗提供了有力的手段。１　Ｇ－ＣＳＦ的生物学作用人类Ｇ－ＣＳＦ由单个基因编码,其基因位于１７号染色体的ｑ２１－２２区,长约２．５ｋｂ,有５个外显子和４个内含子;其…     

肝癌患者血清可溶性肿瘤坏死因子受体I的表达及临床意义

用ＥＬＩＳＡ方法测定了３０例肝癌患者血清可溶性肿瘤坏死因子受体Ｉ（ｓＴＮＦＲ－Ｉ）水平。结果表明，肝癌患者血清ｓＴＮＦＲ－Ｉ水平明显高于正常人（ｐ〈０．０１），且血清ｓＴＮＦＲＩ水平与临床分期相关。治疗有效者，血清ｓＴＮＦＲ－Ｉ水平均明显下降（ｐ〈０．０１）。治疗后２￣８月，３例无癌生存者，血清ｓＴＮＦＲ－Ｉ水平降至正常，而７例局部未控或复发转移者，血清ｓＴＮＦＲ－Ｉ水平则进行性上升。提示，血清ｓ     

受体酪氨酸激酶RON与肿瘤

RON的激活与人体内某些上皮源性肿瘤的发生存在密切关系.在体内,RON基因主要表达于上皮细胞及食管、胃、小肠、肾脏、宫颈、皮肤、膀胱等组织和器官,在肺泡、甲状腺、心脏、平滑肌、成纤维细胞、内皮细胞中几乎没有表达.RON在其配体巨噬细胞刺激蛋白的刺激下,激活后具有调节上皮细胞扩散、转移、微管形成等功能.同时,RON本身也可以诱导细胞发生上皮一间质转化,引起细胞扩散和浸润.研究RON在肿瘤发生中的作用可以为其作为靶向药物干预治疗提供可靠的依据.     

甲状腺功能亢进症患者外周血中粒细胞集落刺激因子及其受体表达的意义

目的:观察甲状腺功能亢进(简称甲亢)患者外周血中粒细胞集落刺激因子(G-CSF)和其受体(G-CS-FR)的表达及临床意义。方法:采用双抗体夹心ELISA法和G-CSFR单克隆抗体及流式细胞技术,检测63例初发甲亢患者、22例应用抗甲状腺药物(ATD)治疗控制的甲亢患者及43例正常人外周血中G-CSF和G-CSFR的表达水平,分析粒细胞计数与G-CSF水平及G-CSFR表达的关系。结果:G-CSFR表达率、白细胞总数及中性粒细胞绝对值三组间均无明显差异(P>0.05)。初发甲亢组和甲亢控制组血清G-CSF水平均明显高于正常对照组(P<0.01和P<0.05);各组中G-CSF和G-CSFR与白细胞计数之间无明显的相关性(P>0.05)。结论:甲亢患者外周血中G-CSF水平是升高的,可能与机体免疫反应有关。     

粒细胞集落刺激因子受体在急性髓性白血病骨髓CD34+细胞中的表达及其临床意义研究

目的 评价粒细胞集落刺激因子(G-CSF)应用于急性髓性白血病(AML)化疗患者的安全性.方法 对2008年3月至2009年1月天津医科大学总医院血液肿瘤科62例AML患者[初治或复发AML33例,完全缓解(CR)AML 29例]和16名正常人采用流式细胞术(FCM)和半定量逆转录一聚合酶链反应(RT-PCR)检测骨髓CD34+细胞G-CSFR的表达.结果 CD114+CD34+/CD34+比值在初治或复发组、CR组及对照组分别为(11.69±2.91)%、(31.84±8.62)%、(32.87±8.44)%(P<0.05),CR组和对照组比较差异无统计学意义(P>0.05).G.CSF受体mRNA表达在初治或复发组、CR组及对照组分别为(30.52±6.21)%、(85.13±21.25)%、(91.57±18.64)%(P<0.05),CR组和对照组比较差异无统计学意义(P>0.05).动态随访了13例AML患者,初治及CR时骨髓CD34+细胞上CD114+CD34+/CD34+比值分别为(12.58±2.00)%、(30.13±7.09)%,初治时低于CR时(P<0.05).G-CSF受体mRNA表达分别为(32.23±5.69)%、(83.27±19.79)%,初治组低于CR组(P<0.05).结论 G-CSF受体在AML患者骨髓CD34+细胞上的表达低于正常人CD34+细胞上的表达,故G-CSF用于治疗AML化疗后粒细胞缺乏期是安全的.     

肝癌生长抑素受体mRNA表达的研究

生长抑素（SST）是一种广泛存在于人体内分泌系统中环状的多肽类激素，近来SST及其类似物对肝癌诊断及治疗方面的作用正日益受到关注，其多种生物活性的发挥是通过靶细胞膜上的特异性受体介导的。本实验应用逆转录聚合酶链反应（RTPCR）检测原发性及转移性肝癌癌组织、癌旁组织、非癌肝组织、人肝癌细胞株HepG2及人肝细胞QSG-7701中生长抑素受体（SSTR）五种亚型mRNA的表达情况。     

粒细胞集落刺激因子受体内含子基因态性与系统性红斑狼疮的关系

目的探讨粒细胞集落刺激因子受体（G—CSFR）内含子基因多态性的分布特点及与系统性红斑狼疮（SEE）易感性和临床特征的关系。方法采用聚合酶链反应-单链构象多态性（PCR—SSCP）分析方法检测144例SEE患者和489名正常对照组G—CSFR基因第5内含子9606位点及第12内含子14233位点基因型。结果SEE患者G—CSFR基因第5内含子9606位点AG基因型频率（61．8％）及G等位基因频率（37．8％）较对照组（47．0％及27．6％）明显升高（P〈0．01）；而AA基因型频率（31．3％）及A等位基因频率（62．2％）较对照组（48．9％及72．4％）明显降低（P〈0．01）。SEE伴临床表现及实验室指标异常的患者与不伴临床表现及实验室检查异常的SEE患者相比较，各基因型频率及各等位基因频率差异无统计学意义（P均〉0．05）。在全部病例和对照组中未发现G-CSFR基因第12内含子14233位点的单核苷酸多态现象。结论SEE患者G-CSFR基因第5内含子9606位点AG基因型及G等位基因与SEE发病相关，各基因型及等位基因与SEE各临床表现及各项实验室指标无关。G-CSFR基因可能是SEE易感基因。     

雄激素受体与肝细胞肝癌预后的关系

目的　探讨雄激素受体（ＡＲ） 与肝细胞肝癌（ ＨＣＣ） 的关系。方法　用葡聚糖包裹活性炭饱和吸附法检测４７例ＨＣＣ 癌组织和癌旁肝组织中ＡＲ 的含量。对４３ 例病人进行术后随访９ ～６０ 个月。结果　３３ 例ＡＲ 阳性（３－０ ～１００ｆｍｏｌ／ｍｇ 蛋白） ,１４ 例ＡＲ 阴性。ＡＲ 阳性病人与ＡＲ 阴性病人的一、五年存活率没有统计学差别（ Ｐ＞ ０－０５） ,三年存活率ＡＲ 阳性者（３５－５ ％ ） 显著低于ＡＲ 阴性者（６６－７ ％ ）（ Ｐ＜ ０－０５） 。复发时间ＡＲ 阳性者（２１－４ ±１５－２ 月） 与ＡＲ 阴性者（３３－９±１３－９ 月） 有显著差异（ Ｐ＜ ０－０５） 。结论　雄激素能促进ＨＣＣ 的增长及对周围组织的侵袭,部分ＨＣＣ 具有雄激素依赖性,ＡＲ 可作为ＨＣＣ 的预后指标之一     

巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径和细胞内胆固醇酯积聚的影响

为探索巨噬细胞集落刺激因子、清道夫受体、氧化型低密度脂蛋白与动脉粥样硬化的关系，观察了重组人巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径的影响以及重组人巨噬细胞集落刺激因子对氧化型低密度脂蛋白所致细胞内胆固醇酯积聚的影响．结果发现重组人巨噬细胞集落刺激因子能增加培养的小鼠腹腔巨噬细胞表面的清道夫受体数目，使之对氧化型低密度脂蛋白的结合和降解呈现剂量和时间依赖性增加，并使细胞内胆固醇酯积聚增多．表明巨噬细胞集落刺激因子可通过增加清道夫受体数目使小鼠腹腔巨噬细胞对氧化型低密度脂蛋白的结合和降解增多，从而增加细胞内胆固醇酯含量，促进动脉壁内泡沫细胞形成．     

血清AFP联合GP73在原发性肝癌的诊断中的意义

目的探讨甲胎蛋白（AFP）和高尔基体糖蛋白-73（GP73）在原发性肝癌患者和健康者血清中的水平及其诊断价值。方法回顾性分析原发性肝癌患者的类型及可能的危险因素,采用电化学发光法及酶联免疫法（ELISA）分别对37例肝癌患者和38例健康体验者进行血清AFP及GP73的含量检测,分析AFP、GP73及AFP＋GP73在肝癌诊断中的敏感性和特异性。结果 37例原发性肝癌患者中,34例（91.89%）为肝细胞癌,28例（75.68%）有病毒性肝炎病史。肝癌患者血清中AFP和GP73的含量分别为（418.31±189.93）ng/ml和（252.03±238.34）ng/ml,显著高于对照组的（16.28±9.39）ng/ml和（34.03±22.67）ng/ml,均P〈0.01。AFP和GP73联合检测肝癌的敏感性及特异性可达86.49%和84.20%,与单项检测相比,可明显提高诊断的准确性。结论血清中AFP及GP73含量是原发性肝癌发生、发展的重要检测指标,二者联合检测对肝癌患者诊断具有重要意义。     

Macrophage-colony-stimulating factor (CSF-1) induces proliferation, chemotaxis, and reversible monocytic differentiation in myeloid progenitor cells transfected with the human c-fms/CSF-1 receptor cDNA.

The c-fms protooncogene encodes the receptor for macrophage-colony-stimulating factor (CSF-1). Expression vectors containing either normal or oncogenic point-mutated human c-fms genes were transfected into interleukin 3 (IL-3)-dependent 32D cells in order to determine the effects of CSF-1 signaling in this murine clonal myeloid progenitor cell line. CSF-1 was shown to trigger proliferation in association with monocytic differentiation of the 32D-c-fms cells. Monocytic differentiation was reversible upon removal of CSF-1, implying that CSF-1 was required for maintenance of the monocyte phenotype but was not sufficient to induce an irrevocable commitment to differentiation. Human CSF-1 was also shown to be a potent chemoattractant for 32D-c-fms cells, suggesting that CSF-1 may serve to recruit monocytes from the circulation to tissue sites of inflammation or injury. Although c-fms did not release 32D cells from factor dependence, point-mutated c-fms[S301,F969] (Leu-301----Ser, Tyr-969----Phe) was able to abrogate their IL-3 requirement and induce tumorigenicity. IL-3-independent 32D-c-fms[S301,F969] cells also displayed a mature monocyte phenotype, implying that differentiation did not interfere with progression of these cells to the malignant state. All of these findings demonstrate that a single growth factor receptor can specifically couple with multiple intracellular signaling pathways and play a critical role in modulating cell proliferation, differentiation, and migration.     

Therapeutic applications of macrophage colony-stimulating factor-1 (CSF-1) and antagonists of CSF-1 receptor (CSF-1R) signaling

Macrophage-colony stimulating factor (CSF-1) signaling through its receptor (CSF-1R) promotes the differentiation of myeloid progenitors into heterogeneous populations of monocytes, macrophages, dendritic cells, and bone-resorbing osteoclasts. In the periphery, CSF-1 regulates the migration, proliferation, function, and survival of macrophages, which function at multiple levels within the innate and adaptive immune systems. Macrophage populations elicited by CSF-1 are associated with, and exacerbate, a broad spectrum of pathologies, including cancer, inflammation, and bone disease. Conversely, macrophages can also contribute to immunosuppression, disease resolution, and tissue repair. Recombinant CSF-1, antibodies against the ligand and the receptor, and specific inhibitors of CSF-1R kinase activity have been each been tested in a range of animal models and in some cases, in patients. This review examines the potential clinical uses of modulators of the CSF-1/CSF-1R system. We conclude that CSF-1 promotes a resident-type macrophage phenotype. As a treatment, CSF-1 has therapeutic potential in tissue repair. Conversely, inhibition of CSF-1R is unlikely to be effective in inflammatory disease but may have utility in cancer.     

Monoclonal antibodies to the human CSF-1 receptor (c-fms proto-oncogene product) detect epitopes on normal mononuclear phagocytes and on human myeloid leukemic blast cells

The first monoclonal antibodies (MoAbs) to epitopes in the extracellular domain of the human c-fms proto-oncogene product (receptor for the macrophage colony stimulating factor, CSF-1) were used with flow cytometric techniques to study receptor expression on normal human peripheral blood monocytes, bone marrow cells, and leukemic blasts. On normal cells CSF-1 receptors were restricted in their expression to cells of the mononuclear phagocyte lineage. CSF-1 receptors were detected on leukemic blasts from 15 (30%) of 50 children with acute myeloid leukemia, compared with four (15%) of 26 adults. By contrast, detectable CSF-1 receptors were uniformly absent on blasts from 19 children with acute lymphoblastic leukemia. CSF-1 receptors on normal monocytes and myeloid leukemia cells could be induced to downmodulate by incubation with either human recombinant CSF-1 or phorbol esters, confirming that the receptors had functional ligand- binding sites and responded to transmodulation by inducers of protein kinase C. The numbers of receptors per cell and the percentage of positive cases were highest for leukemic blasts with cytochemical and morphological features of monocytes. However, CSF-1 receptors were also detected on a subset of leukemic blast cells with features of granulocytic differentiation (FAB subtypes M1 through M3). Southern blotting analyses of DNA from 47 cases of acute myeloid leukemia demonstrated no rearrangements within the 32 kb of genomic sequences that contain CSF-1 receptor coding exons or in the 50 kb upstream of the first coding exon. Analysis of the upstream region of the c-fms locus revealed that sequences representing the terminal 112 untranslated nucleotides of c-fms mRNA map 26 kb 5' to the first coding exon, suggesting that at least one c-fms promoter is separated from the receptor coding sequences by a very long intron. Whereas expression of the CSF-1 receptor in myeloid leukemic blasts is not restricted to cells with monocytic characteristics, the apparently aberrant pattern of receptor synthesis in a subset of cases with granulocytic features appears not to be due to chromosomal rearrangements within 50 kb upstream of sequences encoding the receptor.     

Incomplete restoration of colony-stimulating factor 1 (CSF-1) function in CSF-1-deficient Csf1op/Csf1op mice by transgenic expression of cell surface CSF-1

The primary macrophage growth factor, colony-stimulating factor 1 (CSF-1), is expressed as a secreted glycoprotein or proteoglycan found in the circulation or as a biologically active cell surface glycoprotein (csCSF-1). To investigate the in vivo roles of csCSF-1, we created mice that exclusively express csCSF-1, in a normal tissue-specific and developmental manner, by transgenic expression of csCSF-1 in the CSF-1-deficient osteopetrotic (Csf1(op)/Csf1(op)) background. The gross defects of Csf1(op)/Csf1(op) mice, including growth retardation, failure of tooth eruption, and abnormal male and female reproductive functions were corrected. Macrophage densities in perinatal liver, bladder, sublinguinal salivary gland, kidney cortex, dermis, and synovial membrane were completely restored, whereas only partial or no restoration was achieved in adult liver, adrenal gland, kidney medulla, spleen, peritoneal cavity, and intestine. Residual osteopetrosis, significantly delayed trabecular bone resorption in the subepiphyseal region of the long bone, and incomplete correction of the hematologic abnormalities in the peripheral blood, bone marrow, and spleens of CSF-1-deficient mice were also found in mice exclusively expressing csCSF-1. These data suggest that although csCSF-1 alone is able to normalize several aspects of development in Csf1(op)/Csf1(op) mice, it cannot fully restore in vivo CSF-1 function, which requires the presence of the secreted glycoprotein and/or proteoglycan forms.     

Temporal expression and location of colony-stimulating factor 1 (CSF-1) and its receptor in the female reproductive tract are consistent with CSF-1-regulated placental development.

During pregnancy the mouse uterine epithelial synthesis of the mononuclear phagocyte growth factor designated colony-stimulating factor 1 (CSF-1) is regulated by female sex steroids. To study the role of CSF-1 in the pregnant female reproductive tract, the temporal expression and cellular sites of synthesis of CSF-1 and CSF-1 receptor (CSF-1R) mRNA were determined. CSF-1 mRNA, predominantly the 2.3-kilobase (kb) form, was first detected by in situ hybridization in uterine epithelium prior to implantation on day 3 and subsequently increased, reaching a peak at days 14-15. Its expression was restricted to the uterine epithelium at all stages of gestation and was not localized to areas of implantation. CSF-1R mRNA was first detected in maternal decidua at day 6. It was expressed in the decidua basalis during placentation, after which its expression declined. At day 7.5, trophectodermal cells also expressed CSF-1R mRNA; during placentation, it was found also in the diploid trophoblasts. The high level of CSF-1R mRNA expression by trophoblast giant cells was independent of their location around the conceptus. There was a differential distribution of CSF-1R mRNA expression in the mature placenta, with expression in the giant trophoblastic layer greater than spongiotrophoblastic layer greater than labyrinthine layer until term. Yolk sac cells also expressed low levels of CSF-1R mRNA. The coincidence of uterine CSF-1 mRNA expression and CSF-1 synthesis with both placental growth and CSF-1R mRNA expression in decidual cells and trophoblasts strongly implicates CSF-1 in the regulation of placental growth and differentiation.     

Colony-stimulating factor 1-mediated regulation of a chimeric c-fms/v-fms receptor containing the v-fms-encoded tyrosine kinase domain.

A chimeric gene specifying the 308 N-terminal amino acids of the extracellular ligand binding domain of the human c-fms protooncogene joined to the remainder of the feline v-fms oncogene product encodes a functional colony-stimulating factor 1 (CSF-1) receptor. When expressed in mouse NIH 3T3 fibroblasts, the chimeric gene product was rapidly transported to the cell surface, was autophosphorylated on tyrosine only in response to human recombinant CSF-1, underwent ligand-induced but not phorbol ester-induced down-modulation, and stimulated CSF-1-dependent cell proliferation. By contrast, the C-terminally truncated glycoprotein encoded by the v-fms oncogene is partially inhibited in its transport to the plasma membrane, is constitutively phosphorylated on tyrosine, and is relatively refractory to both ligand-induced and phorbol ester-induced down-modulation. Although the v-fms oncogene can transform cells in the absence of CSF-1, its tyrosine kinase activity and turnover can be appropriately regulated by the human c-fms-encoded ligand binding domain. The results confirm that C-terminal truncation of the c-fms gene is insufficient to activate its transforming potential and suggest that an additional mutation in its distal extracellular domain is required for oncogenic activation.     

Inhibition of colony-stimulating factor-1 activity by monoclonal antibodies to the human CSF-1 receptor

Four of 12 monoclonal antibodies (MoAbs) directed to different epitopes in the extracellular domain of the human colony-stimulating factor-1 receptor (CSF-1R, the c-fms proto-oncogene product) specifically inhibit CSF-1 binding to receptor-bearing cells. All four antibodies abrogated CSF-1-dependent colony formation by human bone marrow-derived macrophage precursors and by mouse NIH-3T3 cells expressing a transduced human c-fms gene. In addition, one of these antibodies (designated MoAb 2-4A5) interfered with the ligand-independent proliferation of NIH-3T3 cells transformed by an oncogenic, mutant c- fms allele. Unlike CSF-1 itself, neither MoAb 2-4A5 nor the other three inhibitory antibodies (MoAbs 12-2D6, 12-3A1, and 12-3A3) induced CSF-1R internalization or degradation. These antibodies should prove useful not only for identifying and quantitating CSF-1R on receptor-bearing cells but for abrogating specific receptor signals that govern the proliferation and survival of human mononuclear phagocytes.