Physiologic role of signal transaction pathways in primary cultured human erythroid progenitor cells.

原代培养的人红系祖细胞中信号处理途径的生理作用。

基本信息

批准号：
12670970
负责人：
SAWADA Kenichi
金额：
$ 2.5万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670970/
关键词：
Fas SCF EPO apoptosis ZNF317 TPO FKHRL1 Src GPA c-kit

项目摘要

Development of erythrocytes is a comlex process governed by multiple cytokines. Colony assay revealed the physiologic importance of each of the cytokines. However, biochemical studies of highly purified human colony forming unit-erythroid (CFU-E) generated in vitro from CD34+ cells have only recently begun. Studies from our groups and others suggested that signal transduction in the primary erythroid cells may differ considerably from that in cell lines or the primary cells from other species. We have developed a method to generate purified erythroid progenitors, which are expanded from CD34+ cells in human peripheral blood in a sufficient number to allow for examination of early signaling by EPO or apoptosis. Using these cells, we have found that SCF protected purified human glycophorin A-positive (GPA^+)/c-kit^+ cells from apoptosis, and suggeswted that mediated Src kinase activation is involved in Akt activation and cell survival. Further, we have suggested that SCF prevents Fas-med … More iated apoptosis of erythroid progenitor cells in a manner dependent on the activity of Src-family tyrosine kinases. We also identified active Lyn in erythroid cells. These data suggest the presence of a novel Src-family/AKT dependent function of SCF in the development of erythrocytes. At the same time, we identified a novel Kruppel associated box (KRAB) zinc finger gene, ZNF317. ZNF317 encoded a protein comprised of thirteen Kruppel-like zinc fingers and a KRAB domain. RT-PCR analysis revealed four alternatively splicing products (ZNF317-1 through ZNF317-4). The ZNF317 gene has seven exons and is located in human chromosome 19p13. ZNF317-1 and ZNF317-2 transcripts were ubiquitously expressed, whereas ZNF317-3 and ZNF317-4 transcripts were detected only in lymphocytes, spleen and lung. The expression of ZNF317 mRNA significantly decreased during erythroid maturation. In lymphocytes, ZNF317 expression was reduced in response to mitogenic stimulation. We propose that ZNF317 may play an important role in erythroid maturation and lymphoid proliferation. Less

红细胞的发育是一个受多种细胞因子控制的复杂过程，但对体外 CD34+ 细胞产生的高度纯化的人红细胞集落形成单位 (CFU-E) 的生化研究揭示了每种细胞因子的生理重要性。我们小组和其他人的研究表明，原代红系细胞中的信号转导可能与细胞系或其他物种的原代细胞中的信号转导有很大不同。我们开发了一种产生纯化红系细胞的方法。祖细胞，从人外周血中的 CD34+ 细胞中扩增出足够数量的细胞，以便检查 EPO 或细胞凋亡的早期信号传导，使用这些细胞，我们发现 SCF 可以保护纯化的人血型糖蛋白 A 阳性 (GPA^+)/。 c-kit^+ 细胞免于凋亡，并表明介导的 Src 激酶激活参与 Akt 激活和细胞存活。此外，我们还表明 SCF 可以防止 Fas-med 介导的细胞凋亡。我们还鉴定了红细胞中的活性 Lyn，这些数据表明 SCF 在红细胞发育中存在新的 Src 家族/AKT 依赖性功能。同时，我们鉴定了一种新的 Kruppel 相关盒 (KRAB) 锌指基因 ZNF317，其编码由 13 个 Kruppel 样锌指和组成的蛋白质。 RT-PCR 分析揭示了四个选择性剪接产物（ZNF317-1 至 ZNF317-4），ZNF317 基因具有七个外显子，位于人类染色体 19p13 中，ZNF317-1 和 ZNF317-2 转录本普遍表达。 ZNF317-3 和 ZNF317-4 转录物仅在淋巴细胞、脾脏和肺中检测到。在红细胞成熟过程中，ZNF317 的表达显着降低。在淋巴细胞中，ZNF317 的表达因促有丝分裂刺激而减少。