Establishment of erythroleukemic GAK14 cells and characterization of GATA1 N-terminal domain
Establishment of erythroleukemic GAK14 cells and characterization of GATA1 N-terminal domain
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红白血病 GAK14 细胞的建立和 GATA1 N 末端结构域的表征
DOI：
10.1111/gtc.12084
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发表时间：
2013
期刊：
影响因子：
2.1
通讯作者：
Yamamoto M,
中科院分区：
文献类型：
--
作者：
Mukai HY;Suzuki M;Nagano M;Ohmori S;Otsuki A;Tsuchida K;Moriguchi T;Ohneda K;Shimizu R;Ohneda O;Yamamoto M,
GATA 1是红细胞生成和巨核细胞生成所必需的转录因子。Gata 1基因敲低的杂合子雌性小鼠（Gata1G1.05/+）自发发生成红细胞白血病。在本研究中，我们从Gata1G1.05/+小鼠的白血病细胞中产生了一种新的Gata 1敲低的成红细胞系，命名为GAK 14。尽管在促红细胞生成素和干细胞因子存在下，GAK 14细胞在OP 9基质细胞上保持未成熟表型，但当与DAS 104 - 8饲养细胞共培养时，细胞产生Gr-1-、Mac 1-、B220-、CD 3e-或CD 49 b-阳性造血细胞。然而，GAK 14细胞不产生红系和巨核系，可能是由于缺乏GATA 1。事实上，当与全长GATA 1互补并与胎肝来源的FLS 5基质细胞共培养时，GAK 14细胞能够分化为成熟的红系细胞。当缺乏N-末端结构域的GATA 1被补充时，这种分化潜力受损。已知N末端结构域有助于与唐氏综合征相关的短暂性骨髓生成异常和急性巨核细胞白血病的发病机制。因此，这些结果表明，GAK 14细胞将作为一个强大的工具，用于解剖结构域功能的GATA 1和GATA 1 N末端结构域是必不可少的红系分化的GAK 14细胞。
GATA1 is a transcription factor essential for erythropoiesis and megakaryopoiesis. It has been found thatGata1gene knockdown heterozygous female (Gata1G1.05/+) mice spontaneously develop erythroblastic leukemias. In this study, we have generated a novelGata1knockdown erythroblastic cell line, designated GAK14, from the leukemia cells in theGata1G1.05/+mice. Although GAK14 cells maintain immature phenotype on OP9 stromal cells in the presence of erythropoietin and stem cell factor, the cells produce Gr‐1‐, Mac1‐, B220‐, CD3e‐ or CD49b‐positive hematopoietic cells when co‐cultured with DAS104‐8 feeder cells. However, GAK14 cells did not produce erythroid and megakaryocytic lineages, perhaps due to the absence of GATA1. Indeed, GAK14 cells became capable of differentiating into mature erythroid cells when complemented with full‐length GATA1 and co‐cultured with fetal liver–derived FLS5 stromal cells. This differentiation potential was impaired when GATA1 lacking the N‐terminal domain was complemented. The N‐terminal domain is known to contribute to the pathogenesis of transient abnormal myelopoiesis and acute megakaryoblastic leukemia related to Down syndrome. These results thus showed that GAK14 cells will serve as a powerful tool for dissecting domain function of GATA1 and that the GATA1 N‐terminal domain is essential for the erythroid differentiation of GAK14 cells.