Mapping gene expression patterns during myeloid differentiation using the EML hematopoietic progenitor cell line.
OBJECTIVE
The detailed examination of the molecular events that control the early stages of myeloid differentiation has been hampered by the relative scarcity of hematopoietic stem cells and the lack of suitable cell line models. In this study, we examined the expression of several myeloid and nonmyeloid genes in the murine EML hematopoietic stem cell line.


METHODS
Expression patterns for 19 different genes were examined by Northern blotting and RT-PCR in RNA samples from EML, a variety of other immortalized cell lines, and purified murine hematopoietic stem cells. Representational difference analysis (RDA) was performed to identify differentially expressed genes in EML.


RESULTS
Expression patterns of genes encoding transcription factors (four members of the C/EBP family, GATA-1, GATA-2, PU.1, CBFbeta, SCL, and c-myb) in EML were examined and were consistent with the proposed functions of these proteins in hematopoietic differentiation. Expression levels of three markers of terminal myeloid differentiation (neutrophil elastase, proteinase 3, and Mac-1) were highest in EML cells at the later stages of differentiation. In a search for genes that were differentially expressed in EML cells during myeloid differentiation, six cDNAs were isolated. These included three known genes (lysozyme, histidine decarboxylase, and tryptophan hydroxylase) and three novel genes.


CONCLUSIONS
Expression patterns of known genes in differentiating EML cells accurately reflected their expected expression patterns based on previous studies. The identification of three novel genes, two of which encode proteins that may act as regulators of hematopoietic differentiation, suggests that EML is a useful model system for the molecular analysis of hematopoietic differentiation.
Department of Cell Biology & Biochemistry, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
Yang
Y
Du
Janee L
JL
Campbell
Demet
D
Nalbant
Hyewon
H
Youn
Ann C Hughes
AC
Bass
Everardo
E
Cobos
Schickwann
S
Tsai
Jonathan R
JR
Keller
Simon C
SC
Williams
D016475:3T3 Cells / Q000166:cytology / Q000378:metabolism; D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D015415:Biomarkers; D002460:Cell Line / Q000166:cytology* / Q000378:metabolism; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D004798:Enzymes / Q000096:biosynthesis / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D007377:Interleukin-3 / Q000494:pharmacology; D007668:Kidney / Q000166:cytology; D008647:Mesocricetus; D051379:Mice; D022423:Myeloid Cells / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D018168:Receptors, Retinoic Acid / Q000187:drug effects / Q000235:genetics; D000072482:Retinoic Acid Receptor alpha; D013382:Subtraction Technique; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism
D000943:Antigens, Differentiation; D015415:Biomarkers; D018076:DNA, Complementary; D004798:Enzymes; D007377:Interleukin-3; D012333:RNA, Messenger; C000607072:Rara protein, mouse; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha; D014157:Transcription Factors; D014212:Tretinoin