scholarly journals Peptide nucleic acids targeting β-globin mRNAs selectively inhibit hemoglobin production in murine erythroleukemia cells

2014 ◽  
Vol 35 (1) ◽  
pp. 51-58 ◽  
Author(s):  
GIULIA MONTAGNER ◽  
CHIARA GEMMO ◽  
ENRICA FABBRI ◽  
ALEX MANICARDI ◽  
IGEA ACCARDO ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Peptide Nucleic Acids ◽  
Erythroleukemia Cells ◽  
Murine Erythroleukemia ◽  
Hemoglobin Production ◽  
Murine Erythroleukemia Cells

scholarly journals α-1,6-Fucosyltransferase (FUT8) Inhibits Hemoglobin Production during Differentiation of Murine and K562 Human Erythroleukemia Cells

2013 ◽  
Vol 288 (23) ◽  
pp. 16839-16847 ◽  
Author(s):  
Hitoshi Sasaki ◽  
Takanori Toda ◽  
Toru Furukawa ◽  
Yuki Mawatari ◽  
Rika Takaesu ◽  
...  
Keyword(s):  
Expression Profiles ◽  
K562 Cells ◽  
Erythroid Differentiation ◽  
Cytokine Signaling ◽  
Erythroleukemia Cells ◽  
Transcription Factor Genes ◽  
Murine Erythroleukemia ◽  
Hemoglobin Production ◽  
Core Fucosylation ◽  
Murine Erythroleukemia Cells

Erythropoiesis results from a complex combination of the expression of several transcription factor genes and cytokine signaling. However, the overall view of erythroid differentiation remains unclear. First, we screened for erythroid differentiation-related genes by comparing the expression profiles of high differentiation-inducible and low differentiation-inducible murine erythroleukemia cells. We identified that overexpression of α-1,6-fucosyltransferase (Fut8) inhibits hemoglobin production. FUT8 catalyzes the transfer of a fucose residue to N-linked oligosaccharides on glycoproteins via an α-1,6 linkage, leading to core fucosylation in mammals. Expression of Fut8 was down-regulated during chemically induced differentiation of murine erythroleukemia cells. Additionally, expression of Fut8 was positively regulated by c-Myc and c-Myb, which are known as suppressors of erythroid differentiation. Second, we found that FUT8 is the only fucosyltransferase family member that inhibits hemoglobin production. Functional analysis of FUT8 revealed that the donor substrate-binding domain and a flexible loop play essential roles in inhibition of hemoglobin production. This result clearly demonstrates that core fucosylation inhibits hemoglobin production. Third, FUT8 also inhibited hemoglobin production of human erythroleukemia K562 cells. Finally, a short hairpin RNA study showed that FUT8 down-regulation induced hemoglobin production and increase of transferrin receptor/glycophorin A-positive cells in human erythroleukemia K562 cells. Our findings define FUT8 as a novel factor for hemoglobin production and demonstrate that core fucosylation plays an important role in erythroid differentiation.

Cell cycle analysis of p53-induced cell death in murine erythroleukemia cells

1993 ◽  
Vol 13 (1) ◽  
pp. 711-719
Author(s):  
J J Ryan ◽  
R Danish ◽  
C A Gottlieb ◽  
M F Clarke
Keyword(s):  
Cell Death ◽  
G1 Arrest ◽  
Wild Type ◽  
Transfected Cells ◽  
Endogenous Expression ◽  
Erythroleukemia Cells ◽  
Wild Type P53 ◽  
G1 Cells ◽  
Murine Erythroleukemia ◽  
Murine Erythroleukemia Cells

A temperature-sensitive mutant of murine p53 (p53Val-135) was transfected by electroporation into murine erythroleukemia cells (DP16-1) lacking endogenous expression of p53. While the transfected cells grew normally in the presence of mutant p53 (37.5 degrees C), wild-type p53 (32.5 degrees C) was associated with a rapid loss of cell viability. Genomic DNA extracted at 32.5 degrees C was seen to be fragmented into a characteristic ladder consistent with cell death due to apoptosis. Following synchronization by density arrest, transfected cells released into G1 at 32.5 degrees C were found to lose viability more rapidly than did randomly growing cultures. Following release into G1, cells became irreversibly committed to cell death after 4 h at 32.5 degrees C. Commitment to cell death correlated with the first appearance of fragmented DNA. Synchronized cells allowed to pass out of G1 prior to being placed at 32.5 degrees C continued to cycle until subsequently arrested in G1; loss of viability occurred following G1 arrest. In contrast to cells in G1, cells cultured at 32.5 degrees C for prolonged periods during S phase and G2/M, and then returned to 37.5 degrees C, did not become committed to cell death. G1 arrest at 37.5 degrees C, utilizing either mimosine or isoleucine deprivation, does not lead to rapid cell death. Upon transfer to 32.5 degrees C, these G1 synchronized cell populations quickly lost viability. Cells that were kept density arrested at 32.5 degrees C (G0) lost viability at a much slower rate than did cells released into G1. Taken together, these results indicate that wild-type p53 induces cell death in murine erythroleukemia cells and that this effect occurs predominantly in the G1 phase of actively cycling cells.

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