Hemoglobin Expression in Clones of K562 Cell Line

Testa, Ugo; Vainchenker, William; Beuzard, Yves; Rouyer-Fessard, P; Guerrasio, Angelo; Titeux, M; Lapotre, Philippe; Bouguet, J; Breton-Gorius, J; Rosa, J.

doi:10.1111/j.1432-1033.1982.tb05835.x

Cited by 19 publications

(10 citation statements)

References 41 publications

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“…Among them, 190 miRNAs were less expressed in CB compared to AB reticulocytes. To reduce this list of 190 miRNAs, we employed an artificial in vitro system: we isolated a clone of the myelogenous leukemia cell line K562 that did not express HbF at detectable levels, whereas HbF increased to almost 50% of the expressed hemoglobin after treatment with hemin [27], [28], [29]. However, the Philadelphia chromosome (Ph1)-positive K562 cells cannot be compared to any normal stage of erythropoiesis, neither before nor after hemin treatment [30].…”

Section: Resultsmentioning

confidence: 99%

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MicroRNA-96 Directly Inhibits γ-Globin Expression in Human Erythropoiesis

et al. 2011

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Fetal hemoglobin, HbF (α2γ2), is the main hemoglobin synthesized up to birth, but it subsequently declines and adult hemoglobin, HbA (α2β2), becomes predominant. Several studies have indicated that expression of the HbF subunit γ-globin might be regulated post-transcriptionally. This could be confered by ∼22-nucleotide long microRNAs that associate with argonaute proteins to specifically target γ-globin mRNAs and inhibit protein expression. Indeed, applying immunopurifications, we found that γ-globin mRNA was associated with argonaute 2 isolated from reticulocytes that contain low levels of HbF (<1%), whereas association was significantly lower in reticulocytes with high levels of HbF (90%). Comparing microRNA expression in reticulocytes from cord blood and adult blood, we identified several miRNAs that were preferentially expressed in adults, among them miRNA-96. The overexpression of microRNA-96 in human ex vivo erythropoiesis decreased γ-globin expression by 50%, whereas the knock-down of endogenous microRNA-96 increased γ-globin expression by 20%. Moreover, luciferase reporter assays showed that microRNA-96 negatively regulates expression of γ-globin in HEK293 cells, which depends on a seedless but highly complementary target site located within the coding sequence of γ-globin. Based on these results we conclude that microRNA-96 directly suppresses γ-globin expression and thus contributes to HbF regulation.

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Section: Resultsmentioning

confidence: 99%

“…HbF induction in K562 cells was performed, as described [27], [28], [29] with minor modifications, by adding 50 µM of hemin (Sigma, Buchs, Switzerland) to the medium for two days.…”

Section: Methodsmentioning

confidence: 99%

MicroRNA-96 Directly Inhibits γ-Globin Expression in Human Erythropoiesis

et al. 2011

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“…One possible explanation for the different levels of mRNA would be the cloning process, as it has been shown that cloning of K562 can result in different subclones with a high heterogeneity in their ability of haemoglobin synthesis and accumulation patterns (Testa et al, 1982). Analysis of the endogenous and hybrid globin gene mRNA levels and DNA blot analysis shows that: firstly, the cells containing the human hybrid eand -y-gene sequences transcribed the genes and processed the mRNA correctly.…”

Section: Discussionmentioning

confidence: 99%

Differential expression of human globin genes introduced in K562 cells.

Kioussis¹,

Wilson²,

Khazaie³

et al. 1985

The EMBO Journal

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We have constructed a series of hybrid globin genes between the 5′ half of the human epsilon‐, gamma‐ and beta‐globin genes, and the 3′ half of the rabbit beta‐globin gene. These hybrid genes were introduced into the cell line K562. Analysis of the hybrid mRNA populations show that only the hybrid epsilon‐ and gamma‐globin genes are expressed in these cells in concordance with the endogenous K562 globin gene expression. It is therefore likely that these cells contain transcriptional factors which allow the specific expression of introduced epsilon‐ and gamma‐, but not beta‐globin genes, irrespective of their chromosomal localization.

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“…Culture of human K562 cell line and analysis of cell differentiation The human K562 cell line was cultured as previously reported (Testa et al , 1982). Cell cultures were initiated at the concentration of 2 × 10 5 cells/ml in 75 cm 3 flasks and collected at different times for analysis.…”

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confidence: 99%

Resveratrol, a natural dietary phytoalexin, possesses similar properties to hydroxyurea towards erythroid differentiation

et al. 2001

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Summary. Resveratrol, a natural dietary polyphenol, has been postulated to be implicated in the cardioprotective effect of red wine and the low incidence of breast and prostate cancers among vegetarians and Orientals respectively. This compound inhibits ribonucleotide reductase as does hydroxyurea, the first therapeutic agent used in the treatment of sickle cell disease. Using the human erythroleukaemic K562 cell line as an in vitro model, we show here that 50 mmol/l of resveratrol induced a higher haemoglobin production (sevenfold) in K562 cells than 500 mmol/l of hydroxyurea (3´5-fold). This erythroid differentiation was linked to a dose-and time-dependent inhibition of cell proliferation associated with an equivalent increased expression of p21 mRNA, but with a higher increased level of p21 protein (sixfold) for cells treated with resveratrol than for those treated with hydroxyurea (1´5-fold). We also show that 50 mmol/l of resveratrol and 25 mmol/l of hydroxyurea induced variable but similar inhancements of fetal haemoglobin synthesis in cultured erythroid progenitors for the majority of the sickle cell patients studied. These inductions were linked to, but not correlated with, a variable decrease in erythroid burst-forming unit clone number. Taken together, these results show that resveratrol merits further investigations in sickle cell disease therapy.

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Hemoglobin Expression in Clones of K562 Cell Line

Cited by 19 publications

References 41 publications

MicroRNA-96 Directly Inhibits γ-Globin Expression in Human Erythropoiesis

MicroRNA-96 Directly Inhibits γ-Globin Expression in Human Erythropoiesis

Differential expression of human globin genes introduced in K562 cells.

Resveratrol, a natural dietary phytoalexin, possesses similar properties to hydroxyurea towards erythroid differentiation

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