HUMAN LEUKEMIC DIFFERENTIATION--ANTI-SENSE RNA STUDIES

可诱导的人髓系白血病细胞系为实验提供 获取白血病细胞分化和正常的事件 造血术。有可能细胞分化策略 最终可以提供基于以下内容的替代方案 用于人类临床处理的细胞破坏 恶性血液病。反义RNA转染法 分析提供了一种直接评估功能角色的方法 某些基因，例如原癌基因和集落刺激 因子(CSF)基因，在白血病分化计划中。近期 我们实验室的研究已经确定了高拷贝的用处 有效反义RNA的上体复制子个数 人造血细胞基因表达的介导性抑制 细胞。所提出的实验目标是：1)优化 基于表型复制子的组成性和诱导性转基因 在人造血细胞中的表达，2)研究 原癌基因和脑脊液mRNA在一组诱导性细胞中的表达 人髓系白血病细胞系(HL-60、ML-1、K562、KG-1、 U937)，并特别注意年表和谱系 基因表达变化的特异性，3)反义RNA 突变分析以确定哪些(如果有的话)表达了 原癌基因和CSF在白血病中起重要作用 4)1，25-二羟基维生素D3的衍生化 (1，25(OH)2D3)诱导特异性HL-60cDNA通过 差异筛选和克隆方法及其功能 反义RNA转染法对这些基因的鉴定 实验。拟议的研究应有助于 对正常和非正常的分子机制的理解 白血病的造血功能，并可提供对 反义RNA技术在肿瘤研究中的有效应用 特异性基因在人类造血细胞功能和功能中的作用 差异化。

Inducible human myeloid leukemic cell lines provide experimental access to events in both leukemic cell differentiation and normal hematopoiesis. It is possible that cytodifferentiation strategies could ultimately provide alternatives to ones based upon cytodestruction for the clinical management of human hematopoietic malignancies. Anti-sense RNA transfection analysis offers a means for directly assessing the functional role of certain genes, e.g., protooncogenes and colony stimulating factor (CSF) genes, in leukemic differentiation programs. Recent studies in our laboratory have established the utility of high copy number episomal replicons for the effective anti-sense RNA- mediated inhibition of gene expression in human hematopoietic cells. The proposed experiments are directed at: 1) optimization of episomal replicon-based constitutive and inducible transgene expression in human hematopoietic cells, 2) investigation of protooncogene and CSF mRNA expression in a panel of inducible human myeloid leukemic cell lines (HL-60, ML-1, K562, KG-1, U937), with specific attention to the chronology and lineage specificity of changes in gene expression, 3) anti-sense RNA mutational analysis to determine which, if any, expressed protooncogenes and CSFs play an obligatory role in leukemic differentiation, and 4) derivation of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) induction-specific HL-60 cDNAs through differential screening and cloning methodologies and functional evaluation of these genes through anti-sense RNA transfection experiments. The proposed studies should contribute to an understanding of the molecular mechanisms underlying normal and leukemic hematopoiesis, and could provide general insights into the effective use of anti-sense RNA technology in studying the role of specific genes in human hematopoietic cell function and differentiation.