MECHANISM OF 1-25-D3 ANTI-PROLIFERATIVE ACTION IN CML

1-25-D3 在 CML 中的抗增殖作用机制

• 批准号: 3195105
• 负责人: STEPHEN R LASKY
• 金额: $ 13.66万
• 依托单位: ROGER WILLIAMS HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-07 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3195105
• 关键词: 1,25 dihydroxycholecalciferol; DNA binding protein; antileukemic agent; gel electrophoresis; molecular cloning; mutant; myelogenous leukemia; neoplastic cell; protein kinase; receptor binding; receptor mediated endocytosis; surface antigens

RWLeu-4 in a recently established, Philadelphia chromosome positive (Ph1) cell line isolated from a patient in the "blast phase" of chronic myelogenous leukemia (CML). Treatment of RWLeu-4 with la,25(OH)2 Vitami D3 (1,25-D3), phorbol esters (PMA), or DMSO causes these cells to cease proliferating and to differentiate into cells with phenotypic, functional, and cell surface antigenic characteristics of monocytes and/or macrophages. We have recently derived a variant of this cell line, designated RD3, that in resistant to the anti-proliferative effect of 1,25-D3 but in still induced to differentiate in response to these agents. Using both the wild type and the resistant cells, we will investigate the mechanism of anti-proliferative action of 1,25-D3 on these CML cells. We propose to characterize the 1,25-D3 receptors in bc RWLeu-4 and RD3 cells using DNA-cellulose elution profiles, partial peptide maps, and DNA-cellulose binding profiles. If differences are found, we will further characterize the genes coding for the receptors using molecular cloning techniques. If no differences are found we will examine changes in other signal trasduction systems such as protein kinase C and other protein kinases, phosphatidylinositol metabolism interferon, cyclic nucleotide production and changes in the phospholipid, content of the cell membrane. Exploiting differences in gene expression in RWLeu-4 and RD3 in response to 1,25-D3 as detected by differential hybridization of cDNA's made from treated and untreated rwleu4 and rd3 cells, we will investigate the factors specific for the regulation of proliferation per se. By doing so, we hope to further understand the processes that prevent normal maturation of cells in CML and begin to elucidate the factors that regulate cellular proliferation. Furthermore these studies will enhance our understanding of the mechanism of action of 1,25-Da in general, and regulation of proliferation and maturation in hematopoietic cells in specific.

RWLEU-4在最近建立的费城染色体阳性中 （PH1）在慢性的“爆炸阶段”中从患者中分离出的细胞系 脊髓性白血病（CML）。用LA的RWLEU-4处理25（OH）2 Vitami D3（1,25-D3），Phorbol酯（PMA）或DMSO导致这些细胞停止 增殖并通过表型分化为细胞 单核细胞的功能和细胞表面抗原特性 和/或巨噬细胞。我们最近得出了该单元的一种变体 线，指定的RD3，具有抗抗增殖作用的能力 1,25-d3，但仍引起响应的响应 代理商。使用野生型和抗性细胞，我们将 研究1,25-d3对抗增殖作用的机制 这些CML细胞。我们建议表征卑诗省的1,25-d3受体 RWLEU-4和RD3细胞使用DNA-纤维素洗脱轮廓，部分 肽图和DNA-纤维素结合曲线。如果有差异 发现，我们将进一步表征编码受体的基因 使用分子克隆技术。如果找不到差异，我们 将检查其他信号示威系统（例如蛋白质）的变化 激酶C和其他蛋白激酶，磷脂酰肌醇代谢 干扰素，环状核苷酸的产生和变化 磷脂，细胞膜的含量。利用差异 RWLEU-4和RD3中的基因表达响应于1,25-D3 由处理和未经处理的cDNA的差异杂交 RWLEU4和RD3细胞，我们将研究特定的因素 本质的调节。通过这样做，我们希望进一步 了解防止CML细胞正常成熟的过程 并开始阐明调节细胞增殖的因素。 此外，这些研究将增强我们对 作用机理 一般的1,25-DA，并调节增殖和成熟 特异性造血细胞。