ROLE OF ETS1 IN LYMPHOID CELLS

ETS1 在淋巴细胞中的作用

• 批准号: 3774907
• 负责人: T S PAPAS
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3774907
• 关键词: B lymphocyte; DNA binding protein; T lymphocyte; cell differentiation; cell growth regulation; cellular oncology; clone cells; electroporation; erythroid stem cell; flow cytometry; gene expression; genetic enhancer element; genetic promoter element; leukocyte activation /transformation; lymphatic tissue; lymphokines; messenger RNA; mitogens; myeloid stem cell; northern blottings; oncogenes; oncoproteins; phorbols; surface antigens; transcription factor

The ets family of genes encodes transcription factors, binds to purine- rich motifs (A/CGGAAGT/C), and regulates promoter/enhancer activities of cellular and viral genes. Members of the ets family include: ETS1, ETS2, ERG, ELK, ERGB/FLI, PU.1/SpiI, ELF1, GABPalpha, PEA3, ER81, and ER71 genes. To understand the function of the ets family of genes during hematopoietic differentiation, well-characterized cell lines representing erythroid, myeloid, and lymphoid lineages have been used to study ets expression by RNA blot analyses. The expression of PEA3 and ERG was higher in the myeloid precursor cell lineage than in functionally-mature myeloid cells. The GABPalpha, ETS2 mRNAs are expressed in most of the cell lines studied. However, ETS1 expression was much higher in lymphoid cells than in cells of myeloid or erythroid origin. ERGB, GABPalpha, GABPbeta and ETS2 was also expressed in thymocytes and splenic B- and T- cells. During T-cell activation by mitogens, both ETS1 and ERGB expression was reduced. Stimulation of promyelocytic cell line HL60 by phorbol esters, GABPalpha and beta mRNAs is induced during an early phase of differentiation. Therefore, it appears that ets gene products may play a role in cell growth, differentiation, and functional maturation of hematopoietic cells. To elucidate the role of the ets family of genes in T-cells, eukaryotic expression vectors containing the DNA-binding domain and variant forms of ETS1 were transfected into Jurkat T-cells by electro- poration, and independent clones were selected in G418. Characterization of these clones for expression of exogenous ets gene products, and their effect on target genes, is in progress. In particular, FACS and RNA blot analysis reflecting the expression pattern of surface antigen and growth properties of the transfected cells (growth rate, tumorigenesis), the production of lymphokines (RNA and ELISA), and the modulation of activities of specific candidate genes targeted by the ets family of genes (RNA blots, CAT assays, differential display), are being performed.

ets家族基因编码转录因子，结合嘌呤- 丰富的基序（A/CGGAAGT/C），并调节启动子/增强子的活性， 细胞和病毒基因。 ets家族的成员包括：ETS 1，ETS 2， ERG、ELK、ERGB/FLI、PU.1/SpiI、ELF1、GABPalpha、PEA3、ER 81和ER 71 基因. 为了了解ets家族基因在 造血分化，充分表征的细胞系代表 红系、髓系和淋巴系已被用于研究ets 通过RNA印迹分析表达。 PEA 3和ERG的表达呈负相关。 在髓样前体细胞谱系中比在功能成熟的 骨髓细胞 GABPalpha、ETS 2 mRNA在大多数细胞中表达 研究的细胞系。 然而，ETS 1在淋巴结中的表达要高得多， 细胞比髓系或红系起源的细胞中的细胞更少。 ERGB、GABPalpha、 GABA β和ETS 2也在胸腺细胞和脾B-和T-细胞中表达。 细胞 在有丝分裂原激活T细胞期间，ETS 1和ERGB 表达减少。 人早幼粒细胞系HL 60的刺激作用 佛波酯、GABA α和β mRNA在早期阶段被诱导 的差异。 因此，看来ets基因产物可能发挥 在细胞生长、分化和功能成熟中的作用 造血细胞 阐明ets家族基因在 T细胞，含有DNA结合结构域的真核表达载体 和ETS 1的变体形式通过电转染进入Jurkat T细胞， 穿孔，并在G418中选择独立的克隆。 表征 表达外源性ets基因产物的克隆，以及它们的 对靶基因的影响正在进行中。 特别是，FACS和RNA印迹 反映表面抗原和生长的表达模式的分析 转染细胞的特性（生长速率，肿瘤发生）， 产生淋巴因子（RNA和ELISA），以及调节 由基因的ets家族靶向的特定候选基因的活性 (RNA印迹、CAT测定、差异显示）。