Erythroid differentiation of clonal Rauscher erythroleukemia cells in response to erythropoietin or dimethyl sulfoxide

Science ◽  
1980 ◽  
Vol 210 (4465) ◽  
pp. 74-76 ◽  
Author(s):  
A. Sytkowski ◽  
A. Salvado ◽  
G. Smith ◽  
C. McIntyre ◽  
N. deBoth
Keyword(s):  
Dimethyl Sulfoxide ◽  
Erythroid Differentiation ◽  
Erythroleukemia Cells

scholarly journals Induction and inhibition of Friend erythroleukemia cell differentiation by pyrimidine analogs: analysis of the requirement for intracellular accumulation and incorporation into DNA.

1982 ◽  
Vol 2 (8) ◽  
pp. 1020-1024 ◽  
Author(s):  
J A Bilello ◽  
K K Gauri ◽  
J Kühne ◽  
G Warnecke ◽  
G Koch
Keyword(s):  
Cell Differentiation ◽  
Dimethyl Sulfoxide ◽  
Dna Analysis ◽  
Erythroid Differentiation ◽  
Intracellular Accumulation ◽  
Induced Differentiation ◽  
Alkyl Chains ◽  
Erythroleukemia Cells ◽  
Erythroleukemia Cell ◽  
Pyrimidine Analogs

Alkyldeoxyuridines which differ from thymidine by a C5 substitution of straight or branched alkyl chains of two to six carbon atoms have been tested for their ability to be taken up, phosphorylated, and incorporated into DNA. Analysis of the uptake of 5-ethyl-2'-deoxyuridine and 5-propyl-2'-deoxyuridine (n-PrdU)--similar to both thymidine and 5-bromo-2'-deoxyuridine--indicates that transport is dependent upon a functional cellular thymidine kinase. All of the aforementioned pyrimidines with the exception of n-PrdU are phosphorylated to the triphosphate and incorporated into DNA. The homologs 5-iso-propyl-2'-deoxyuridine (iso-PrdU) and 5-hexyl-2'-deoxyuridine are neither transported into the cell, phosphorylated, nor incorporated into DNA. These analogs were tested (i) for their ability to induce in the absence of dimethyl sulfoxide and (ii) to determine whether they enhance or inhibit dimethyl sulfoxide-induced differentiation of Friend erythroleukemia cells. Inhibition of erythroid differentiation appears to require the incorporation of thymidine analogs into DNA, and thus only 5-ethyl-2'-deoxyuridine and 5-bromo-2'-deoxyuridine were effective in inhibiting dimethyl sulfoxide-induced differentiation. The observation that iso-PrdU, and to a lesser extent n-PrdU and 5-propyldeoxyuridine monophosphate, induce differentiation under conditions in which they are not detectable intracellularly is strong evidence that this class of inducer acts at the cell membrane.

scholarly journals Role of the PU.1 transcription factor in controlling differentiation of Friend erythroleukemia cells.

1992 ◽  
Vol 12 (7) ◽  
pp. 2967-2975 ◽  
Author(s):  
S Schuetze ◽  
R Paul ◽  
B C Gliniak ◽  
D Kabat
Keyword(s):  
Transcription Factor ◽  
Dimethyl Sulfoxide ◽  
Nuclear Localization ◽  
Erythroid Differentiation ◽  
Polyclonal Antiserum ◽  
Cellular Genes ◽  
Erythroleukemia Cells ◽  
Chemically Induced ◽  
Preleukemic Stage

Both viral and cellular genes have been directly implicated in pathogenesis of Friend viral erythroleukemia. The virus-encoded gp55 glycoprotein binds to erythropoietin receptors to cause mitogenesis and differentiation of erythroblasts. However, if the provirus integrates adjacent to the gene for the PU.1 transcription factor, the cell loses its commitment to terminally differentiate and becomes immortal, as indicated by its transplantability and by its potential for indefinite growth in culture (C. Spiro, B. Gliniak, and D. Kabat, J. Virol. 63:4434-4437, 1989; R. Paul, S. Schuetze, S. L. Kozak, and D. Kabat, J. Virol. 65:464-467, 1991). To test the implications of these results, we produced polyclonal antiserum to bacterially synthesized PU.1, and we used it to analyze PU.1 expression throughout leukemic progression and during chemically induced differentiation of Friend erythroleukemia (F-MEL) cell lines. This antiserum identified three electrophoretically distinct PU.1 components in extracts of F-MEL cells and demonstrated their nuclear localization. Although PU.1 proteins are abundant in F-MEL cells, they are absent or present in only trace amounts in normal erythroblasts or in differentiating erythroblasts from the preleukemic stage of Friend disease. Furthermore, chemicals (dimethyl sulfoxide or N,N'-hexamethylenebisacetamide) that overcome the blocked differentiation of F-MEL cells induce rapid declines of PU.1 mRNA and PU.1 proteins. The elimination of PU.1 proteins coincides with recommitment to the program of erythroid differentiation and with loss of immortality. These results support the hypothesis that PU.1 interferes with the commitment of erythroblasts to differentiate and that chemicals that reduce PU.1 expression reinstate the erythropoietic program.

scholarly journals Regulation of beta-globin mRNA accumulation by heme in dimethyl sulfoxide (DMSO)-sensitive and DMSO-resistant murine erythroleukemia cells

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1662-1667 ◽  
Author(s):  
Y Fukuda ◽  
H Fujita ◽  
L Garbaczewski ◽  
S Sassa
Keyword(s):  
Dimethyl Sulfoxide ◽  
Erythroid Differentiation ◽  
Mrna Levels ◽  
Beta Globin ◽  
Globin Mrna ◽  
Mrna Accumulation ◽  
Erythroleukemia Cells ◽  
Dmso Treatment ◽  
Free Heme ◽  
Murine Erythroleukemia

The level of mRNA encoding beta-globin was examined in dimethyl sulfoxide (DMSO)-sensitive (DS), and DMSO-resistant (DR) murine erythroleukemia (MEL) cells. DR cells lack erythroid-specific delta- aminolevulinate (ALA) synthase (AL-AS-E), and fail to undergo erythroid differentiation following treatment with DMSO. Treatment of cells with DMSO markedly increased ALAS-E mRNA in DS cells, while the same treatment downregulated the nonspecific ALA synthase (ALAS-N) mRNA levels in both DS and DR cells. The levels of beta-globin mRNA, heme content, and hemoglobin in DS cells increased, while those in DR cells decreased following treatment with DMSO. Treatment of DR cells with hemin caused an increase in beta-globin mRNA and hemoglobin, and partially restored the DMSO-mediated suppression of beta-globin mRNA and hemoglobin synthesis. DMSO treatment decreased heme oxygenase (HO) mRNA in hemin-treated DS cells, but not in hemin-treated DR cells. These findings indicate that heme is necessary for accumulation of the beta-globin transcript during erythroid differentiation, and that hemin- mediated HO induction becomes markedly downregulated in differentiated erythroid cells, presumably because less free heme is available for HO induction by a greater demand for the synthesis of hemoglobin.

scholarly journals Regulation of beta-globin mRNA accumulation by heme in dimethyl sulfoxide (DMSO)-sensitive and DMSO-resistant murine erythroleukemia cells

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1662-1667 ◽  
Author(s):  
Y Fukuda ◽  
H Fujita ◽  
L Garbaczewski ◽  
S Sassa
Keyword(s):  
Dimethyl Sulfoxide ◽  
Erythroid Differentiation ◽  
Mrna Levels ◽  
Beta Globin ◽  
Globin Mrna ◽  
Mrna Accumulation ◽  
Erythroleukemia Cells ◽  
Dmso Treatment ◽  
Free Heme ◽  
Murine Erythroleukemia

Abstract The level of mRNA encoding beta-globin was examined in dimethyl sulfoxide (DMSO)-sensitive (DS), and DMSO-resistant (DR) murine erythroleukemia (MEL) cells. DR cells lack erythroid-specific delta- aminolevulinate (ALA) synthase (AL-AS-E), and fail to undergo erythroid differentiation following treatment with DMSO. Treatment of cells with DMSO markedly increased ALAS-E mRNA in DS cells, while the same treatment downregulated the nonspecific ALA synthase (ALAS-N) mRNA levels in both DS and DR cells. The levels of beta-globin mRNA, heme content, and hemoglobin in DS cells increased, while those in DR cells decreased following treatment with DMSO. Treatment of DR cells with hemin caused an increase in beta-globin mRNA and hemoglobin, and partially restored the DMSO-mediated suppression of beta-globin mRNA and hemoglobin synthesis. DMSO treatment decreased heme oxygenase (HO) mRNA in hemin-treated DS cells, but not in hemin-treated DR cells. These findings indicate that heme is necessary for accumulation of the beta-globin transcript during erythroid differentiation, and that hemin- mediated HO induction becomes markedly downregulated in differentiated erythroid cells, presumably because less free heme is available for HO induction by a greater demand for the synthesis of hemoglobin.

Induction and inhibition of Friend erythroleukemia cell differentiation by pyrimidine analogs: analysis of the requirement for intracellular accumulation and incorporation into DNA

1982 ◽  
Vol 2 (8) ◽  
pp. 1020-1024
Author(s):  
J A Bilello ◽  
K K Gauri ◽  
J Kühne ◽  
G Warnecke ◽  
G Koch
Keyword(s):  
Cell Differentiation ◽  
Dimethyl Sulfoxide ◽  
Dna Analysis ◽  
Erythroid Differentiation ◽  
Intracellular Accumulation ◽  
Induced Differentiation ◽  
Alkyl Chains ◽  
Erythroleukemia Cells ◽  
Erythroleukemia Cell ◽  
Pyrimidine Analogs

Alkyldeoxyuridines which differ from thymidine by a C5 substitution of straight or branched alkyl chains of two to six carbon atoms have been tested for their ability to be taken up, phosphorylated, and incorporated into DNA. Analysis of the uptake of 5-ethyl-2'-deoxyuridine and 5-propyl-2'-deoxyuridine (n-PrdU)--similar to both thymidine and 5-bromo-2'-deoxyuridine--indicates that transport is dependent upon a functional cellular thymidine kinase. All of the aforementioned pyrimidines with the exception of n-PrdU are phosphorylated to the triphosphate and incorporated into DNA. The homologs 5-iso-propyl-2'-deoxyuridine (iso-PrdU) and 5-hexyl-2'-deoxyuridine are neither transported into the cell, phosphorylated, nor incorporated into DNA. These analogs were tested (i) for their ability to induce in the absence of dimethyl sulfoxide and (ii) to determine whether they enhance or inhibit dimethyl sulfoxide-induced differentiation of Friend erythroleukemia cells. Inhibition of erythroid differentiation appears to require the incorporation of thymidine analogs into DNA, and thus only 5-ethyl-2'-deoxyuridine and 5-bromo-2'-deoxyuridine were effective in inhibiting dimethyl sulfoxide-induced differentiation. The observation that iso-PrdU, and to a lesser extent n-PrdU and 5-propyldeoxyuridine monophosphate, induce differentiation under conditions in which they are not detectable intracellularly is strong evidence that this class of inducer acts at the cell membrane.

Role of the PU.1 transcription factor in controlling differentiation of Friend erythroleukemia cells

1992 ◽  
Vol 12 (7) ◽  
pp. 2967-2975
Author(s):  
S Schuetze ◽  
R Paul ◽  
B C Gliniak ◽  
D Kabat
Keyword(s):  
Transcription Factor ◽  
Dimethyl Sulfoxide ◽  
Nuclear Localization ◽  
Erythroid Differentiation ◽  
Polyclonal Antiserum ◽  
Cellular Genes ◽  
Erythroleukemia Cells ◽  
Chemically Induced ◽  
Preleukemic Stage

Both viral and cellular genes have been directly implicated in pathogenesis of Friend viral erythroleukemia. The virus-encoded gp55 glycoprotein binds to erythropoietin receptors to cause mitogenesis and differentiation of erythroblasts. However, if the provirus integrates adjacent to the gene for the PU.1 transcription factor, the cell loses its commitment to terminally differentiate and becomes immortal, as indicated by its transplantability and by its potential for indefinite growth in culture (C. Spiro, B. Gliniak, and D. Kabat, J. Virol. 63:4434-4437, 1989; R. Paul, S. Schuetze, S. L. Kozak, and D. Kabat, J. Virol. 65:464-467, 1991). To test the implications of these results, we produced polyclonal antiserum to bacterially synthesized PU.1, and we used it to analyze PU.1 expression throughout leukemic progression and during chemically induced differentiation of Friend erythroleukemia (F-MEL) cell lines. This antiserum identified three electrophoretically distinct PU.1 components in extracts of F-MEL cells and demonstrated their nuclear localization. Although PU.1 proteins are abundant in F-MEL cells, they are absent or present in only trace amounts in normal erythroblasts or in differentiating erythroblasts from the preleukemic stage of Friend disease. Furthermore, chemicals (dimethyl sulfoxide or N,N'-hexamethylenebisacetamide) that overcome the blocked differentiation of F-MEL cells induce rapid declines of PU.1 mRNA and PU.1 proteins. The elimination of PU.1 proteins coincides with recommitment to the program of erythroid differentiation and with loss of immortality. These results support the hypothesis that PU.1 interferes with the commitment of erythroblasts to differentiate and that chemicals that reduce PU.1 expression reinstate the erythropoietic program.

Inhibition of DMSO-induced differentiation by hyperthermia in a murine erythroleukemia cell system

1984 ◽  
Vol 62 (11) ◽  
pp. 1091-1096 ◽  
Author(s):  
G. P. Raaphorst ◽  
E. I. Azzam ◽  
J. Borsa ◽  
M. Einspenner ◽  
J. A. Vadasz
Keyword(s):  
Heat Treatment ◽  
Dimethyl Sulfoxide ◽  
Erythroid Differentiation ◽  
Staining Technique ◽  
Cell System ◽  
Differentiation Process ◽  
Erythroleukemia Cells ◽  
Dmso Treatment ◽  
Erythroleukemia Cell ◽  
Murine Erythroleukemia

Friend erythroleukemia cells were induced by dimethyl sulfoxide (DMSO) into erythroid differentiation, as characterized by the production of hemoglobin. Induction increased with DMSO concentrations up to 1.5% v/v, at which point about 90% of the cell population produced hemoglobin as measured by a benzidine-staining technique. Heat treatment at 39.0–40.5 °C during a 7-day-incubation period, for differentiation in the presence of DMSO, resulted in the inhibition of hemoglobin induction. Also, acute heat treatments at 41.5–46.0 °C before or after the addition of DMSO resulted in the inhibition of DMSO induction. This effect was greatest when DMSO was present during heating. The results support the conclusion that hyperthermia inhibits the differentiation process which is induced by DMSO treatment.

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