ROLE OF ETS1 IN LYMPHOID CELLS

ETS1原癌基因在淋巴系T细胞中高水平表达。 细胞。ETS1的表达与α基因的表达有关 发育过程中的T细胞受体(TCRpha)。ETS1的表达为 维持在成熟的静息T细胞中，其水平在 激活。ETS1是一种转录因子，与一种富含嘌呤的 基序(AGGAAGT)发现于许多不同基因的调节区， 即淋巴因子、TCRα、FOS、ETS1、ETS2。要了解其功能 在T细胞的ETS1基因产物中，ETS1的活性被表达- ING反义ETS1 RNA。反义RNA能够与 正义的RNA，从而阻止蛋白质的形成。这些同源基因 然后，将使用转基因细胞系来研究 发生在ETS1功能通路的下游。为此，Jurkat T- 用有限稀释法亚克隆细胞，用于转染法。这个 全长ETS1 cDNA及其亚片段(代表EST的5‘端区域 C DNA)克隆到基于巨细胞病毒的表达载体和异构体中 表达反义RNA的载体。将该基因导入Jurkat细胞 这些表达载体通过电穿孔。独立的克隆是 在G418中被选中，正在进一步调查中。特别是，FACS 和Northern分析反映了表面抗原的表达模式 (TCRpha、TCRbeta、CD3、CD4)，检测转基因细胞的生长特性 细胞(生长率，肿瘤发生)，淋巴因子的产生 细胞(ELISA法)，以及特定候选基因的调控 ETS1(使用Northern分析、免疫沉淀和CAT检测)是 正在进行一系列全面的实验。

The ETS1 proto-oncogene is expressed at a higher level in lymphoid T- cells. The expression of ETS1 is related to the expression of the alpha T-cell receptor (TCRalpha) during development. The expression of ETS1 is maintained in mature resting T cells, and its level decreases after activation. ETS1 is a transcription factor and binds to a purine-rich motif (AGGAAGT) found in the regulatory regions of many different genes, i.e., lymphokines, TCRalpha, FOS, ETS1, ETS2. To understand the function of ETS1 gene products in T-cells, the ETS1 activity is blocked by express- ing antisense ETS1 RNA. Antisense RNAs are able to form hybrids with sense RNA, thus blocking the formation of proteins. These isogenic transfectant cell lines will then be used to investigate the changes that occur downstream of ETS1 functional pathways. For this purpose, Jurkat T- cells were subcloned by limited dilution and used for transfection. The full-length ETS1 cDNA and subfragments (representing the 5' region of the cDNA) were cloned in cytomegalovirus-based expression vectors and episomal vectors to express antisense RNA. Jurkat cells were transfected with these expression vectors by electroporation. Independent clones were selected in G418 and are under further investigation. In particular, FACS and Northern analysis reflecting the expression pattern of surface antigen (TCRalpha, TCRbeta, CD3, CD4), the growth properties of the transfected cell (growth rate, tumorigenesis), the production of lymphokines by these cells (ELISA), and the modulation of specific candidate genes targeted by ETS1 (using Northern analysis and immunoprecipitation and CAT assays) are being performed in a series of comprehensive experiments.