干细胞因子和受体在神经系统中的表达及其生物学效应

干细胞因子（stem cell factor,SCF)是一种多功能细胞因子,其受体由原癌基因c-kit编码,称为c-kit受体（c-kitR)。SCF－c-kitR不论在胚胎发育期还是成年期的神经系统,均有广泛的表达。体内外大量研究提示：SCF/c0kitR信号系统在神经系统生长、分化过程中具有多种生物学效应,表现对神经嵴4细胞体外分化的影响,对神经胶质细胞（小胶质细胞、星形胶质细胞和少突胶质细胞）的调控作用,并与神经内分泌功能相关。     

可溶性人 SCF 在大肠杆菌中高效表达

干细胞因子(SCF)是一种参与机体生长发育过程中多种细胞生长调控的多功能细胞因子,该因子是 c-kit 原癌基因编码受体的配体,主要由基质细胞产生.研究发现 SCF 在机体内以两种形式存在,一种是表达于细胞膜表面的膜型 SCF,另一种为分泌至细胞外的可溶性SCF,后者相当于膜型 SCF 的膜外活性区域,由164—165个氨基酸(aa)组成,目前国外用于研究的重组 SCF 产品主要指在大肠杆菌或真核系统中表达的分泌型 SCF,此研究首次在国内报道在大肠杆菌中表达这种类型的重组人 SCF.     

癌基因与癌变

原癌基因(proto-oncogene)是细胞关键的调节基因,调控细胞的增殖和分化。在致癌因子的攻击下,proto-onc 可被激活为癌基因c-onc(或v-onc),后者通过其过量或变质的表达产物(癌蛋白)使细胞癌变。本文综述了该领域最近1～3年的进展;着重阐述了细胞原癌基因的激活机理以及癌蛋白的结构和功能。     

细胞增殖的调节和癌基因的作用

细胞周期受到正负两类因子的调节,GF刺激静止细胞开始增殖,已分化的成熟细胞能抑制增殖。虽然影响细胞增殖的因素很多,只有基因及其产物才是决定因素,其中包括在细胞周期一定时相瞬间表达活性的CCD基因。原癌基因除能编码GF、GFR和转导物外,另有c-fos、c-myc、c-ras和p53为CCD基因。原癌基因经过突变、重排和倍增后,导致细胞增殖失控,引起癌变。     

肿瘤发生发展的分子机理(2)

人体内的原癌基因与肿瘤抑制基因编码许多种蛋白控制细胞的生长和增殖,在诱导癌变过程中起关键作用．原癌基因获得显性表达功能和肿瘤抑制基因丧失隐性表达功能的突变是致癌的。人类已经具有了解每一种肿瘤的特征基因改变的能力,并因此可以深入研究肿瘤发生和进展机制,全面改进肿瘤的早期诊断。     

c-myc基因上游转录单位的研究

c-myc是一种与禽类逆转录病毒M-29v-myc同源的原癌基因。已有证据表明,c-myc在B细胞瘤发生和发展过程中起着重要作用。在正常细胞中,c-myc一般保持着低水平的表达,而当细胞发生癌变时,c-myc的表达则增高。然而,在正常细胞中,细胞是如何保     

实验性癫痫时脑内c—fos基因与阿片肽及兴奋性氨基酸的关系

细胞癌基因也称原癌基因,其命名与被确定时所在的逆转录病毒和细胞的名称有关。c-fos是v-fos的相应细胞对应物,V-fos是FJB鼠骨瘤病毒中分离出来的,c-fos的蛋白产物与v-fos相似,仅在羧端有差异,c-fos基因正常情况下不会引起癌变,它与细胞生长、分化有关。近来一些实验进一步证明c-fos还与调节细胞功能有关,许多细胞外刺激能诱导c-fos转录增加,一些能激活钙通道的条件、药物,或神经递质如细胞外高钾、钙通道激活剂BAYK     

抗癌基因及其致癌机理

近年来,分子遗传学技术的广泛应用,推动和促进了肿瘤遗传变异研究的深入发展。有关血液系恶性肿瘤的研究已确认至少有两种涉及原癌基因激活的机理是细胞恶变转化的原因。即:慢性粒细胞性白血病(CML)中c-abl原癌基因被异常激活的质变     

氧化与细胞凋亡

细胞凋亡(apoptosis)是细胞的主动死亡,它参与调节机体许多生理及病理过程．近年的研究表明细胞凋亡与氧化有密切关系．当细胞外诸如辐射、高氧、高温、感染、衰老等信息通过活性氧传入细胞,引起细胞脂质过氧化或与细胞凋亡有关的基因的表达,通过一系列生化反应,最终发生细胞凋亡．原癌基因bcl-2可能起着调节作用．     

九种蝗虫精子发生过程中c-kit蛋白特异性表达比较分析(英文)

c-kit蛋白在昆虫和哺乳动物生殖细胞发育中发挥着重要作用。本研究通过免疫组织化学法和生物统计方法,比较分析了隶属于直翅目蝗总科的9种蝗虫精子发生过程中c-kit蛋白表达差异。结果显示,蝗虫精子发生过程中c-kit蛋白均出现阶段特异性表达,并且在9种蝗虫种间c-kit蛋白表达出现显著差异。     

Primary structure of c-kit: relationship with the CSF-1/PDGF receptor kinase family--oncogenic activation of v-kit involves deletion of extracellular domain and C terminus.

The protein kinase domains of v-kit, the oncogene of the acute transforming feline retrovirus HZ4-FeSV (HZ4-feline sarcoma virus), CSF-1R (macrophage colony stimulating factor receptor) and PDGFR (platelet derived growth factor receptor) display extensive homology. Because of the close structural relationship of v-kit, CSF-1R and PDGFR we predicted that c-kit would encode a protein kinase transmembrane receptor (Besmer et al., 1986a; Yarden et al., 1986). We have now determined the primary structure of murine c-kit from a DNA clone isolated from a brain cDNA library. The nucleotide sequence of the c-kit cDNA predicts a 975 amino acid protein product with a calculated mol. wt of 109.001 kd. It contains an N-terminal signal peptide, a transmembrane domain (residues 519-543) and in the C-terminal half the v-kit homologous sequences (residues 558-925). c-kit therefore contains the features which are characteristic of a transmembrane receptor kinase. Comparison of c-kit, CSF-1R and PDGFR revealed a unique structural relationship of these receptor kinases suggesting a common evolutionary origin. The outer cellular domain of c-kit was shown to be related to the immunoglobulin superfamily. The sites of expression of c-kit in normal tissue predict a function in the brain and in hematopoietic cells. N-terminal sequences which include the extracellular domain and the transmembrane domain as well as 50 amino acids from the C-terminus of c-kit are deleted in v-kit. These structural alterations are likely determinants of the oncogenic activation of v-kit.(ABSTRACT TRUNCATED AT 250 WORDS)     

IL-4 regulates c-kit proto-oncogene product expression in human mast and myeloid progenitor cells.

The c-kit proto-oncogene encodes the receptor for a novel hemopoietic cytokine, termed stem cell factor (SCF) or mast cell growth factor (MGF) according to its stimulating spectrum. The human receptor for SCF/MGF is expressed in a subset of normal bone marrow progenitor cells, in leukemic myeloid cells, and in mast cells. In the present study, the effects of recombinant human growth regulators (IL-1 through -9, granulocyte-macrophage/granulocyte/macrophage-CSF, IFN, and TNF) on c-kit proto-oncogene product expression were analyzed by indirect immunofluorescence, by using the anti-SCF/MGFR mAb YB5.B8, and Northern blot analyses, by using a c-kit oligonucleotide probe. Of all cytokines tested, IL-4 was found to down-regulate expression of YB5.B8 Ag in the human mast cell line HMC-1 (maximum inhibition, 51.05 +/- 16.36% mean fluorescence intensity of control; p less than 0.02), as well as in primary leukemic myeloid cells. IL-4 was also found to down-regulate expression of YB5.B8 Ag in normal enriched bone marrow progenitor cells. The effects of IL-4 on expression of YB8.B8 Ag in myeloid/mast cell progenitors was dose and time dependent (maximum effects observed on days 2 and/or 4, by using 50 U/ml of rIL-4) and could be neutralized by using anti-IL-4 mAb. Moreover, IL-4 was found to down-regulate expression of c-kit mRNA in leukemic myeloid cells as well as in HMC-1 cells. Together, these observations identify IL-4 as a regulator of c-kit proto-oncogene product expression in the human system. The effects of IL-4 on human hemopoietic progenitor cells and mast cells may be mediated in part through regulation of SCF/MGFR expression.     

Dominant negative and loss of function mutations of the c-kit (mast/stem cell growth factor receptor) proto-oncogene in human piebaldism.

Piebaldism is an autosomal dominant disorder of melanocyte development and is characterized by congenital white patches of skin and hair from which melanocytes are completely absent. A similar disorder of the mouse, "dominant white spotting" (W), results from mutations of the c-kit proto-oncogene, which encodes the cellular tyrosine kinase receptor for the mast/stem cell growth factor. We have identified c-kit gene mutations in three patients with piebaldism. A missense substitution (Phe----Leu) at codon 584, within the tyrosine kinase domain, is associated with a severe piebald phenotype, whereas two different frameshifts, within codons 561 and 642, are both associated with a variable and relatively mild piebald phenotype. This is consistent with a possible "dominant negative" effect of missense c-kit polypeptides on the function of the dimeric receptor.     

c-kit protein, a transmembrane kinase: identification in tissues and characterization.

The proto-oncogene c-kit encodes a transmembrane kinase which is related to the receptors for colony-stimulating factor type 1 and platelet-derived growth factor, as well as to the immunoglobulin superfamily. Antibodies specific for the kinase domain of the P80 gag-kit protein of the Hardy-Zuckerman 4 feline sarcoma virus were prepared. These kit-specific antibodies were used to identify and characterize the c-kit protein in cat brain tissue. The c-kit protein product displays an autophosphorylating activity in immune complex kinase assays, and, in turn, this activity was used to identify the c-kit protein in different tissues. In cat brain, a single 145-kilodalton (kDa) glycoprotein was detected. Its N-linked carbohydrates were found to be sensitive to digestion with the endoglycosidases (neuraminidase, endoglycosidase F, and endoglycosidase H), indicating hybrid and/or complex and high-mannose structures. A partial purification of the c-kit protein was achieved by wheat germ agglutinin affinity chromatography, and the autophosphorylating activity of the partially purified c-kit protein was characterized and found to be specific for tyrosine. The kit antibodies cross-react with the murine c-kit protein product, and variant c-kit proteins in different mouse tissues were identified, with sizes of about 145 kDa (brain), 160 kDa (spleen), and 150 kDa (testis).     

The dominant-white spotting (W) locus of the mouse encodes the c-kit proto-oncogene

Mutations at the W locus in the mouse have pleiotropic effects on embryonic development and hematopoiesis. The characteristic phenotype of mutants at this locus, which includes white coat color, sterility, and anemia, can be attributed to the failure of stem cell populations to migrate and/or proliferate effectively during development. Mapping experiments suggest that the c-kit proto-oncogene, which encodes a putative tyrosine kinase receptor, is a candidate for the W locus. We show here that the c-kit gene is disrupted in two spontaneous mutant W alleles, W44 and Wx. Genomic DNA that encodes amino acids 240 to 342 of the c-kit polypeptide is disrupted in W44; the region encoding amino acids 342 to 791 is disrupted in Wx. W44 homozygotes exhibit a marked reduction in levels of c-kit mRNA. These results strongly support the identification of c-kit as the gene product of the W locus.     

The expression of c-kit protein during oogenesis and early embryonic development.

The c-kit proto-oncogene encodes a transmembrane tyrosine kinase receptor and was shown to be allelic with the white-spotting locus (W) of the mouse. Mutations at the W locus have pleiotropic effects on the development of hematopoietic stem cells, melanoblasts, and primordial germ cells. In order to elucidate the role of c-kit protein in gametogenesis and oocyte maturation, we have examined immunohistochemically the expression of c-kit in the ovaries of mice at late fetal and postnatal stages, and in early embryos. By the avidin-biotin-peroxidase (ABC) method using rat anti-mouse c-kit monoclonal antibody, the c-kit protein was detected in ovaries after the time of birth, but not before. The expression of c-kit was observed mainly on the surface of oocytes, but not in granulosa cells nor in interstitial regions. Oocytes of primordial to fully grown Graafian follicles showed the c-kit protein. When ovulation was induced by hCG, the expression of c-kit in ovulated unfertilized oocytes was weaker than in oocytes of Graafian follicles. In 1-cell embryos the c-kit protein was still observed, but with cell division its expression further decreased, and it was not detected in embryos of 4-cell, 8-cell, and morula stages. In summary, the highest expression of c-kit was observed on the surface of oocytes arrested in the diplotene stage of meiotic prophase. With ovulation and the resumption of meiotic maturation, its expression declined. These results suggest that the c-kit protein may play some role in meiotic arrest, oocyte growth, and oocyte maturation.     

Activation of the human c-kit product by ligand-induced dimerization mediates circular actin reorganization and chemotaxis.

The proto-oncogene c-kit is allelic with the murine white spotting (W) locus and encodes a transmembrane protein tyrosine kinase that is structurally related to the receptors for platelet-derived growth factor (PDGF) and colony-stimulating factor-1 (CSF-1). Recently the ligand for the c-kit product, stem cell factor (SCF), was identified in both transmembrane and soluble forms. In order to examine the mechanism for receptor activation by SCF and biological properties of the activated c-kit product, we transfected the wild-type human c-kit cDNA into porcine aortic endothelial cells. We found that the receptor was down-regulated and transmitted a mitogenic signal in response to stimulation with soluble SCF. We also demonstrate that SCF induces dimerization of the c-kit product in intact cells, and that dimerization of the receptor is correlated with activation of its kinase. Activation of the c-kit product by SCF was found to induce circular actin reorganization indistinguishable from that mediated by the PDGF beta-receptor in response to PDGF-BB. Furthermore, soluble SCF was a potent chemotactic agent for cells expressing the c-kit product, a property which might be of importance during embryonic development.