Promotive Effect of C/EBPε Overexpression on Differentiation of Human Myelomonocytic Leukemia Cell Line U-937
10.3321/j.issn:1000-467X.2006.11.009
- VernacularTitle:增强C/EBPε表达对人粒-单核性白血病细胞U-937分化的作用
- Author:
Rong CAI
1
;
Bing-Bing DAI
;
Ke LI
;
Hong-Jie WANG
;
Wei-Rong XU
;
Jia-Min WANG
;
Jian LU
Author Information
1. 上海交通大学
- Keywords:
CCAAT-enhancer binding protein ε;
Cell differentiation;
cMyc;
Human myelomonocytic leukemia;
U-937 cells
- From:Chinese Journal of Cancer
2006;25(11):1368-1373
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND & OBJECTIVE: CCAAT-enhancer binding protein ε (C/EBPε) is a kind of nuclear transcriptional factor expressed predominantly in myeloid cells, and may be a critical regulator of myeloid differentiation, which can activate the transcription of a subset of myeloidspecific genes. This study was to detect the cell-specific expression of C/EBPε, and to investigate the effect of C/EBPε overexpression on c-Myc expression, cell cycle distribution, and cell differentiation of human myelomonocytic leukemia cell line U-937. METHODS: The expression of C/EBPε in 5 hematopoietic cell lines (U-937, NB4, HL-60, K-562 and Jurkat T cell lines) was tested by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). According to the RT-PCR results, human myelomonocytic leukemia cell line U-937 was selected and transfected with pcDNA3.1-C/EBPε32 expression vector by electroperforation, and screened by G418 to yield positive clones which were termed U-937-C/EBPε32. The expression of C/EBPε and c-Myc in U937-C/EBPε32 cells was detected by RT-PCR and Western blot; cell cycle and differentiation of U937-C/EBPε32 cells was analyzed by flow cytometry. RESULTS: C/EBPε overexpression obviously increased the expression of CD11b (a ceil surface marker of granulocyte maturity). The positive rate of CD11b was significantly higher in U-937-C/EBPε32 cells than in U-937 and U-937-pcDNA3.1 cells(92.56% vs.77.46% and 74.81% ), while c-Myc expression and cell cycle had no changes. CONCLUSION: C/EBPε might be an essential transcriptional regulator for U-937 cells differentiating into granulocytes.
