Over-expression of EDAG in the myeloid cell line 32D: induction of GATA-1 expression and erythroid/megakaryocytic phenotype.
Erythroid differentiation-associated gene (EDAG), a hematopoietic tissue-specific transcription regulator, plays a key role in maintaining the homeostasis of hematopoietic lineage commitment. However, the mechanism and genes regulated by EDAG remain unknown. In this study, we showed that overexpression of EDAG in a myeloid cell line 32D led to an erythroid phenotype with increased number of benzidine-positive cells and up-regulation of erythroid specific surface marker TER119. The megakaryocytic specific marker CD61 was also induced significantly. Using a genome-wide microarray analysis and a twofold change cutoff, we identified 332 genes with reduced expression and 288 genes with increased expression. Among up-regulation genes, transcription factor GATA-1 and its target genes including EKLF, NF-E2, Gfi-1b, hemogen, SCL, hemoglobin alpha, beta and megakaryocytic gene GPIX were increased. Silencing of EDAG by RNA interference in K562 cells resulted in down-regulation of these genes. Taken together, EDAG functions as a positive regulator of erythroid/megakaryocytic differentiation in 32D cells associated with the induction of GATA-1 and its target genes.
Ding
Ya-Li
YL
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Xu
Cheng-Wang
CW
Wang
Zhi-Dong
ZD
Zhan
Yi-Qun
YQ
Li
Wei
W
Xu
Wang-Xiang
WX
Yu
Miao
M
Ge
Chang-Hui
CH
Li
Chang-Yan
CY
Yang
Xiao-Ming
XM
D002454:Cell Differentiation; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry; D050982:GATA1 Transcription Factor / Q000235:genetics*; D020868:Gene Silencing; D006801:Humans; D007377:Interleukin-3 / Q000378:metabolism; D020014:K562 Cells; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C435745:HEMGN protein, human; D007377:Interleukin-3; D009687:Nuclear Proteins