Differential expression of multiple unexpected genes during U937 cell and macrophage differentiation detected by suppressive subtractive hybridization.

TitleDifferential expression of multiple unexpected genes during U937 cell and macrophage differentiation detected by suppressive subtractive hybridization.
Publication TypeJournal Article
Year of Publication2000
AuthorsGingras, M-C, Margolin, JF
JournalExp Hematol
Volume28
Issue1
Pagination65-76
Date Published2000 Jan
ISSN0301-472X
KeywordsAcetyltransferases, Alternative Splicing, Antigens, CD, Antigens, CD34, Antigens, Differentiation, Myelomonocytic, Blotting, Northern, Blotting, Southern, Cell Differentiation, Cells, Cultured, Gene Expression, Hematopoietic Stem Cells, Humans, Lipopolysaccharide Receptors, Macrophages, Neoplasm Proteins, Nerve Tissue Proteins, Nucleic Acid Hybridization, Polymerase Chain Reaction, RNA, Messenger, U937 Cells
Abstract

OBJECTIVE: The objective of this study was to identify new markers of myelomonocytic differentiation using a sensitive technique that permits detection of rare differential gene expression.MATERIALS AND METHODS: [corrected] Suppressive subtractive hybridization (SSH) was performed between the human myelomonocytic U937 cell line and 1 alpha, 25-dihydroxyvitamin D3 and transforming growth factor beta 1 differentiated U937 cells. cDNA clones with significant increased expression in differentiated U937 cells over nondifferentiated U937 cells were characterized by sequencing. [corrected] The pattern of differential gene expression obtained by SSH was confirmed by cDNA Southern and Northern blots on the undifferentiated vs. differentiated U937 cells, and by reverse transcriptase polymerase chain reaction on undifferentiated human CD34(+) stem cells isolated from bone marrow vs. peripheral blood CD14(+) mature monocytes.RESULTS: Seven cDNAs never associated with in vitro U937 cell myelomonocytic differentiation (prolactin, 11-beta hydroxysteroid dehydrogenase [11 beta-HSD)] haptoglobin alpha (2FS)-beta precursor, GLIPR, RTVP, the RNA helicase P68, and spermidine-spermine N1-acetyltransferase) were identified. The first five of these genes previously were associated with immune function and the last two are important for intermediary metabolism. Differential expression was confirmed in CD34(+)/CD14(+) monocyte differentiation for all genes but 11 beta-HSD.CONCLUSIONS: We identified six new markers of U937 cell differentiation, which also are differentially expressed during normal human myelomonocytic differentiation.

Alternate JournalExp. Hematol.
PubMed ID10658678
Grant ListK08 CA67495-03 / CA / NCI NIH HHS / United States
R01 CA080200-01 / CA / NCI NIH HHS / United States