Induction of differentiation in Friend erythroleukemia cells with dimethyl sulfoxide, hexamethylene bisacetamide and sodium butyrate is not accompanied by changes in proviral DNA or its expression

Zajac-Kaye, Maria; Brown, Elissa J.; Friend, Charlotte

doi:10.1016/0168-1702(86)90056-0

Cited by 5 publications

(5 citation statements)

References 29 publications

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“…4 ). The data obtained indicate that all the inducers employed, as reported in previous studies by our and other groups 17 , 18 , 33 , 35 , 38 , 39 , 40 , 41 , increased Hb production in K562 cells ( Fig. 4 A).…”

Section: Resultssupporting

confidence: 87%

“…One of the hypotheses derived from the experiments described in this article is that mithramycin might have a direct effect on BCL11A-XL activity. We therefore wanted to further verify whether increases in globin gene expression might be associated with downregulation of BCL11A-XL in other cellular erythroid model systems, such as murine FLCs 38 , 39 and human primary ErPCs isolated from β–thalassemia patients 34 , 35 , 36 . FLCs were cultured for 4 days with 400 nmol/L MTH or 2% DMSO, and the proportion of benzidine-positive cells was determined together with the levels of α-globin, adult β major -globin, embryofetal βh1-globin, and BCL11A mRNAs.…”

Section: Resultsmentioning

confidence: 99%

“…To sustain the concept that the increase in expression of γ-globin genes is inversely associated with BCL11A-XL content, murine FLCs 38 , 39 and ErPCs isolated from β-thalassemia patients were employed, induced by MTH, and analyzed for content of globin and BCL11A-XL mRNAs. Fully in agreement with the conclusions derived from the data on K562(BCL11A-XL) clones, downregulation of BCL11A-XL was observed in FLCs and ErPCs following a MTH-induced increase in globin mRNAs production (β major - and βh1-globin mRNAs in FLCs, γ-globin mRNA in ErPCs).…”

Section: Discussionmentioning

confidence: 99%

“…Human erythroleukemia K562 cells 33 , 34 , 35 , 36 , 37 and murine Friend leukemia cells (FLCs) 38 , 39 were cultured as described 33 , 39 . Cell growth was studied using a Z2 Coulter Counter (Beckman Coulter, Fullerton, CA).…”

Section: Methodsmentioning

confidence: 99%

“…Human erythromyeloblastoid leukemia K562 cells 33 , 34 , 35 , 36 , 37 , K562(BCL11A-XL) clones, and murine Friend leukemia cells (FLCs) 38 , 39 were cultured in a humidified atmosphere of 5% CO 2 /air in RPMI-1640 medium (Gibco Life Technologies, Monza, Italy) supplemented with 10% (v/v) fetal bovine serum (FBS, Biowest, Nuaillé, France), 100 U/mL penicillin, and 100 μg/mL streptomycin. Treatment of cells with erythroid differentiation inducers was carried out by adding the appropriate drug concentrations at the beginning of the cultures [(K562 and K562(BCL11A-XL) clone cells were seeded at 30,000/mL; FLCs were seeded at 70,000/mL)].…”

Section: Supplementary Methodsmentioning

confidence: 99%

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Development and characterization of K562 cell clones expressing BCL11A-XL: Decreased hemoglobin production with fetal hemoglobin inducers and its rescue with mithramycin

Finotti

Gasparello

Breveglieri

et al. 2015

Experimental Hematology

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Induction of fetal hemoglobin (HbF) is considered a promising strategy in the treatment of β-thalassemia, in which production of adult hemoglobin (HbA) is impaired by mutations affecting the β-globin gene. Recent results indicate that B-cell lymphoma/leukemia 11A (BCL11A) is a major repressor of γ-globin gene expression. Therefore, disrupting the binding of the BCL11A transcriptional repressor complex to the γ-globin gene promoter provides a novel approach for inducing expression of the γ-globin genes. To develop a cellular screening system for the identification of BCL11A inhibitors, we produced K562 cell clones with integrated copies of a BCL11A-XL expressing vector. We characterized 12 K562 clones expressing different levels of BCL11A-XL and found that a clear inverse relationship does exist between the levels of BCL11A-XL and the extent of hemoglobinization induced by a panel of HbF inducers. Using mithramycin as an inducer, we found that this molecule was the only HbF inducer efficient in rescuing the ability to differentiate along the erythroid program, even in K562 cell clones expressing high levels of BCL11A-XL, suggesting that BCL11A-XL activity is counteracted by mithramycin.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Supplementary Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Development and characterization of K562 cell clones expressing BCL11A-XL: Decreased hemoglobin production with fetal hemoglobin inducers and its rescue with mithramycin

Finotti

Gasparello

Breveglieri

et al. 2015

Experimental Hematology

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Use of Sodium Butyrate to Enhance Production of Retroviral Vectors Expressing CFTR cDNA

Olsen

Sechelski²

1995

Human Gene Therapy

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Previously, we constructed a retrovirus vector (LCFSN) for transduction and expression of the cDNA encoding the normal human cystic fibrosis transmembrane conductance regulator (CFTR). The titer of virus from amphotropic packaging cells producing the LCFSN vector was low (10(3)-10(4) infectious units/ml). In an attempt to increase virus production, we used sodium butyrate (NaB) to treat murine retrovirus packaging cells producing this vector. NaB treatment increased the production of LCFSN from between 20-fold to greater than 1,000-fold, depending upon the producer clone, thereby resulting in virus titers up to about 1 x 10(7) infectious units/ml. This induction of virus titer could be accounted for, at least in part, by an increase in steady-state levels of full-length vector RNA within the producer cells. With some clonal producer cell lines, lowering the temperature of the virus harvest in combination with NaB treatment resulted in an apparent synergistic increase in virus production. The production of retrovirus vectors containing genes other than CFTR could also be increased by NaB treatment, although the enhancement in titer was modest (2-fold to 10-fold). The increase in virus production was not accompanied by an induction of replication-competent helper virus. NaB treatment also increased the transient production of retroviral vectors following DNA-mediated transfection into packaging cells such that virus titers of greater than 10(6) infectious units/ml could be readily attained.

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Activated macrophages distinguish undifferentiated-tumorigenic from differentiated-nontumorigenic murine erythroleukemia cells

Pak

Fidler

1989

Differentiation

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Induction of differentiation in Friend erythroleukemia cells with dimethyl sulfoxide, hexamethylene bisacetamide and sodium butyrate is not accompanied by changes in proviral DNA or its expression

Cited by 5 publications

References 29 publications

Development and characterization of K562 cell clones expressing BCL11A-XL: Decreased hemoglobin production with fetal hemoglobin inducers and its rescue with mithramycin

Development and characterization of K562 cell clones expressing BCL11A-XL: Decreased hemoglobin production with fetal hemoglobin inducers and its rescue with mithramycin

Use of Sodium Butyrate to Enhance Production of Retroviral Vectors Expressing CFTR cDNA

Activated macrophages distinguish undifferentiated-tumorigenic from differentiated-nontumorigenic murine erythroleukemia cells

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