HUMAN LEUKEMIC DIFFERENTIATION--ANTI-SENSE RNA STUDIES

人类白血病分化--反义RNA研究

• 批准号: 3458970
• 负责人: Mark L Tykocinski
• 金额: $ 9.5万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-05-01 至 1993-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3458970
• 关键词: RNA; biotechnology; cell bank /registry; cell differentiation; colony stimulating factor; complementary DNA; cytogenetics; flow cytometry; gene expression; genetic library; genetic markers; genetic promoter element; genetic transcription; hematopoiesis; immunochemistry; immunofluorescence technique; leukemia; messenger RNA; molecular cloning; molecular oncology; mutant; neoplasm /cancer genetics; neoplastic cell; neoplastic transformation; nucleic acid probes; plasmids; protooncogene; transfection; transposon /insertion element

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.

诱导型人髓系白血病细胞系提供实验 访问白血病细胞分化和正常细胞的事件 造血作用。 细胞分化策略可能 最终可以提供基于的替代方案 细胞破坏用于人类临床管理 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，在白血病分化程序中。 最近的 我们实验室的研究已经证实了高拷贝的实用性 有效反义RNA的附加型复制子数量 介导的人类造血基因表达抑制 细胞。 建议的实验针对：1）优化 基于游离复制子的组成型和诱导型转基因 人类造血细胞中的表达，2) 的研究 一组诱导细胞中原癌基因和 CSF mRNA 表达 人骨髓白血病细胞系（HL-60、ML-1、K562、KG-1、 U937），特别注意年代和血统 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 原癌基因和脑脊液在白血病中发挥着重要作用 分化，以及4) 1,25-二羟基维生素D3的衍生 (1,25(OH)2D3) 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 了解正常和潜在的分子机制 白血病造血，并可以提供一般见解 反义RNA技术在研究中的有效应用 特定基因在人类造血细胞功能中的作用和 差异化。