HUMAN LEUKEMIC DIFFERENTIATION--ANTI-SENSE RNA STUDIES

人类白血病分化--反义RNA研究

• 批准号: 3458972
• 负责人: Mark L Tykocinski
• 金额: $ 8.72万
• 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/3458972
• 关键词: RNA; antisense nucleic acid; 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; replicon; 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.

可诱导的人髓细胞白血病细胞系提供实验 访问白血病细胞分化和正常的事件 造血。 细胞区分化策略可能 最终可以为基于 人类临床管理的细胞降落 造血恶性肿瘤。 抗敏感RNA转染 分析提供了一种直接评估功能角色的手段 某些基因，例如原子基因和菌落刺激 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确定了高拷贝的实用性 有效的抗敏感RNA的数量偶发子复制子 人造血的介导的基因表达抑制作用 细胞。 提出的实验针对：1）优化 基于偶发子复制子的本构和诱导转基因 人类造血细胞中的表达，2）研究 原子量和CSF mRNA表达在一组诱导的面板中 人髓细胞白血病细胞系（HL-60，ML-1，K562，KG-1， U937），特别关注年表和血统 基因表达变化的特异性，3）反义RNA 突变分析以确定哪个（如果有）表示 原子基因和CSF在白血病中起必不可少的作用 分化和4）衍生1,25-二羟基维生素D3 （1,25（OH）2d3）通过 差分筛选和克隆方法和功能 通过抗敏感RNA转染评估这些基因 实验。 拟议的研究应有助于 理解正常正常和的分子机制 白血病造血，可以提供一般的见解 有效利用反义RNA技术在研究 特定基因在人造血细胞功能中的作用和 分化。