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Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells
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Synthesis of erythrocyte-specific proteins in cultured friend leukemia cells

D Kabat, C C Sherton, L H Evans

    Cell
    |July 1, 1975
    PubMed

    Insights

    Dimethyl sulfoxide (Me2SO) induces hemoglobin synthesis in Friend leukemia (FL) cells. These cells exhibit aberrant erythropoiesis, with altered beta-globin chain ratios compared to normal mouse erythrocytes.

    Area of Science:

    • Biochemistry
    • Cell Biology
    • Hematology

    Background:

    • Friend virus-induced erythroleukemia cells (FL cells) provide a model for studying erythropoiesis.
    • Understanding protein synthesis regulation in these cells is crucial for leukemia research.

    Purpose of the Study:

    • To investigate the synthesis of specific proteins, particularly hemoglobin, in dimethyl sulfoxide (Me2SO)-induced FL cells.
    • To compare the globin chain synthesis ratios in induced FL cells with normal mouse erythrocytes.

    Main Methods:

    • Cell culture of Friend line 745 and Ostertag line FSD-1 FL cells.
    • Induction of differentiation using dimethyl sulfoxide (Me2SO).
    • Analysis of protein synthesis, hemoglobin content, globin chain ratios (alpha, betaMajor, betaMinor) via electrophoresis and chromatography.
    • Measurement of carbonic anhydrase activity and detection of 2,3-diphosphoglyceric acid.

    Main Results:

    • Me2SO treatment led to significant hemoglobin synthesis (up to 25% of total protein).
    • FL cells synthesized heme and globin coordinately with a near 1:1 alpha:beta globin ratio.
    • Significant differences in betaMajor:betaMinor globin chain ratios were observed between the two FL cell lines and normal erythrocytes.
    • Induced FL cells showed higher carbonic anhydrase activity and lacked 2,3-diphosphoglyceric acid.

    Conclusions:

    • Me2SO effectively induces hemoglobinization in FL cells.
    • The globin chain synthesis patterns in induced FL cells suggest an aberrant erythropoiesis.
    • These findings contribute to understanding the molecular mechanisms of erythroid differentiation and potential therapeutic targets in leukemia.

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