HUMAN LEUKEMIC DIFFERENTIATION--ANTI-SENSE RNA STUDIES

可诱导的人髓性白血病细胞系提供了实验性的 获得白血病细胞分化和正常 造血 有可能细胞分化策略 最终可以提供替代方案， 用于人类临床管理的细胞破坏 造血系统恶性肿瘤 反义RNA转染 分析提供了一种直接评估功能作用的方法， 某些基因，例如，原癌基因与集落刺激 因子（CSF）基因，在白血病分化程序。 最近 我们实验室的研究已经确立了高拷贝的效用 有效反义RNA的附加型复制子的数量- 人造血细胞中基因表达的介导抑制 细胞 建议的实验是针对：1）优化 基于附加型复制子的组成型和诱导型转基因 在人类造血细胞中的表达，2）研究 原癌基因和CSF mRNA表达在一组可诱导的 人髓性白血病细胞系（HL-60，ML-1，K562，KG-1， U937），特别注意年表和血统 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 原癌基因和CSFs在白血病中起着必要的作用。 分化，和4）1，25-二羟维生素D3的衍生 （1，25（OH）2D 3）诱导特异性HL-60 cDNA， 差异筛选和克隆方法以及功能 通过反义RNA转染评估这些基因 实验 拟议的研究应有助于 了解正常和 白血病造血，并可以提供一般的见解， 反义RNA技术的有效利用， 特定基因在人类造血细胞功能中的作用， 分化

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.