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: 3195103
• 负责人: STEPHEN R LASKY
• 金额: $ 16.26万
• 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/3195103
• 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在新近建立的费城染色体呈阳性 (PH1)从慢性粒细胞白血病“急变期”患者体内分离的细胞系 髓系白血病(CML)。La，25(OH)2维生素A处理RWLeu-4 D3(1，25-D3)、佛波醇酯(PMA)或DMSO可使这些细胞停止生长 增殖并分化为具有表型的细胞 单核细胞的功能性和细胞表面抗原性 和/或巨噬细胞。我们最近得到了这种细胞的一个变种 命名为RD3的抗增殖作用品系 1，25-D3，但仍被诱导分化以响应这些 探员们。利用野生型和抗性细胞，我们将 1，25-D3抗肿瘤细胞增殖作用机制的研究 这些慢性粒细胞白血病细胞。我们建议对BC的1，25-D3受体进行表征 RWLeu-4和RD3细胞的DNA-纤维素洗脱谱，部分 多肽图和DNA-纤维素结合图谱。如果分歧是 发现后，我们将进一步确定编码受体的基因 使用分子克隆技术。如果没有发现差异，我们 将研究其他信号转导系统的变化，如蛋白质 激酶C和其他蛋白激酶、磷脂酰肌醇代谢 干扰素、环核苷酸的产生和细胞的变化 磷脂，细胞膜的含量。利用中的差异 1，25-D3对RWLeu-4和RD3基因表达的影响 经处理和未处理的cDNA差异杂交 Rwleu4和RD3细胞，我们将研究与RWLeu4和RD3细胞 对扩散本身的调控。通过这样做，我们希望进一步 了解慢性粒细胞白血病中阻止细胞正常成熟的过程 并开始阐明调控细胞增殖的因素。 此外，这些研究将加深我们对 作用机制 一般情况下，1，25-Da的作用以及对细胞增殖和成熟的调节 造血细胞具有特异性。