MACROPHAGE GROWTH: ROLE OF COLONY STIMULATING FACTOR

正常小鼠腹腔渗出液的生长和增殖 巨噬细胞（PEM）在体外是严格依赖于巨噬细胞特异性 称为集落刺激因子（CSF-1）的生长因子。 相比之下， 转化的巨噬细胞肿瘤细胞系不需要外源性CSF-1用于 文化的成长。 最近的研究表明， 某些膜结合蛋白质可能参与细胞的调控， 由特定生长因子诱导的增殖。 此外，几个 癌基因产物已被证明在结构上与 特异性生长因子或生长因子受体。 我们建议使用正常的巨噬细胞和已建立的巨噬细胞 具有不同生长表型的品系作为模型来研究 蛋白磷酸化在巨噬细胞增殖调节中的作用。 首先，我们将发展二维凝胶电泳， 放射自显影技术以鉴定CSF-1诱导的和/或自发的 正常巨噬细胞和已建立的巨噬细胞的磷酸化成分 巨噬细胞-单核细胞肿瘤细胞系。 薄层板电泳 技术将被用来表征化学性质的 磷酸化氨基酸。 第二，针对一种 磷酸酪氨酸部分将被开发为探针，沿着“Western 印迹”技术，以便于鉴定和表征 酪氨酸磷酸化蛋白。 我们将研究是否自分泌CSF-1或CSF样的产生， 活性和酪氨酸磷酸化与致癌 在发育的早期阶段，巨噬细胞转化。 我们 将通过肿瘤病毒转化， 骨髓来源的巨噬细胞，以研究这些细胞系是否产生和 分泌自我刺激活性。 我们还将检查和识别 这些细胞系的磷酸化成分。 我们的假设 工作是自分泌生长因子，酪氨酸磷酸化和 配体-受体相互作用在细胞内 骨髓来源的造血细胞的转化和肿瘤发生。

The growth and proliferation of normal murine peritoneal exudate macrophages (PEM) in vitro is strictly dependent on a macrophage-specific growth factor known as colony-stimulating factor (CSF-1). By contrast, transformed macrophage tumor cell lines do not require exogenous CSF-1 for growth in culture. Recent studies show that tyrosine phosphorylation of certain membrane-bound proteins may be involved in the control of cell proliferation induced by specific growth factors. In addition, several oncogene products have been shown to be structurally related to either specific growth factors or growth factor receptors. We propose to use both normal macrophages and established macrophage cell lines with different growth phenotypes as models to investigate the role of protein phosphorylation in the regulation of macrophage proliferation. First, we will develop two dimensional gel electrophoresis and autoradiography techniques to identify the CSF-1 induced and/or spontaneous phosphorylated components from both mormal macrophages and established macrophage-monocyte tumor cell lines. Thin layer plate electrophoresis techniques will be used to characterize the chemical nature of the phosphorylated amino acids. Second, a monoclonal antibody against a phosphotyrosine moiety will be developed as a probe, along with "Western blot" techniques, to facilitate the identification and characterization of tyrosine-phosphorylated-proteins. We will study whether the production of autocrine CSF-1 or CSF-like activity and tyrosine phosphorylation are linked to oncogenic transformation of macrophages during the early phase of development. We will establish, by tumor virus transformation, immortalized mouse bone marrow-derived macrophages to study whether these cell lines produce and secrete autostimulatory activity. We will also examine and identify phosphorylated components from these cell lines. The hypothesis of our work is that autocrine growth factor, tyrosine phosphorylation and ligand-receptor interaction play important roles in the cellular transformation and oncogenesis of bone marrow-derived hematopoietic cells.