Human leukemia K-562 cells: induction of erythroid differentiation by 5-azacytidine

Gambari, Roberto; Senno, Laura del; Barbieri, Raffaella; Viola, Lorenza; Tripodi, Marco; Raschellà, Giuseppe; Fantoni, A

doi:10.1016/0045-6039(84)90033-2

Cited by 64 publications

(57 citation statements)

References 24 publications

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“…Cell Lines and Culture Conditions-Human erythroleukemia K562(S) cells (44) were cultured in a humidified atmosphere at 5% CO 2 in RPMI 1640 (Flow Laboratories) supplemented with 10% fetal bovine serum (CELBIO), 50 units/ml penicillin, and 50 g/ml streptomycin (45,46). Cell growth was studied by determining the cell number/ml after different days of in vitro cell culture (46).…”

Section: Synthetic Oligonucleotides and Peptidementioning

confidence: 99%

Transcription Factor Decoy Molecules Based on a Peptide Nucleic Acid (PNA)-DNA Chimera Mimicking Sp1 Binding Sites

Borgatti¹,

Lampronti²,

Romanelli³

et al. 2003

Journal of Biological Chemistry

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Peptide nucleic acids (PNAs) are DNA-mimicking molecules in which the sugar-phosphate backbone is replaced by a pseudopeptide backbone composed of N-(2-aminoethyl)glycine units. We determined whether double-stranded molecules based on PNAs and PNA-DNA-PNA (PDP) chimeras could be capable of stable interactions with nuclear proteins belonging to the Sp1 transcription factor family and, therefore, could act as decoy reagents able to inhibit molecular interactions between Sp1 and DNA. Since the structure of PNA/PNA hybrids is very different from that of the DNA/DNA double helix, they could theoretically alter the molecular structure of the double-stranded PNA-DNA-PNA chimeras, perturbing interactions with specific transcription factors. We found that PNA-based hybrids do not inhibit Sp1/DNA interactions. In contrast, hybrid molecules based on PNA-DNA-PNA chimeras are very effective decoy molecules, encouraging further experiments focused on the possible use of these molecules for the development of potential agents for a decoy approach in gene therapy. In this respect, the finding that PDPbased decoy molecules are more resistant than DNA/ DNA hybrids to enzymatic degradation appears to be of great interest. Furthermore, their resistance can even be improved after complexation with cationic liposomes to which PDP/PDP chimeras are able to bind by virtue of their internal DNA structure.In vitro transfection of cis elements that decoy against nuclear factors leads to alteration of gene expression and was recently proposed in molecular medicine as a novel tool for the possible therapy of several disorders (1-12). One of the most effective decoy approaches so far described involves nuclear proteins belonging to the NF-B 1 superfamily (7-9, 13-20).Decoy molecules against NF-B inhibit the expression of NF-B regulated genes (MHC complex genes, IL2 receptor ␣, Igk, IL6, ␦ opioid receptor, preprogalanin, adhesion molecule-1) (20). More recently, dumbell DNA decoy elements against NF-B were demonstrated to be active in inhibiting ex vivo transcription driven by the long terminal repeat (LTR) of human immunodeficiency type-1 virus (HIV-1) (19). In addition to proteins belonging to the NF-B superfamily, decoy molecules for other target transcription factors, such as HNF-1, RFX1, nuclear factor YB, E2F, cAMP-response element, and Sp1, were found to be effective (1-6, 10 -12, 21).As to the molecular targets for the decoy approach, proteins belonging to the Sp1 family are of great interest since these transcription factors are involved in the regulation of the expression of several genes relevant to human pathologies, including those encoding vascular endothelial growth factor, plasminogen-activator inhibitor type-1 (PAI-1), COL1A2, urokinase-type plasminogen activator (uPA), and uPA receptor (3,(21)(22)(23)(24)(25). Sp1 binding sites are also present in the HIV-1 LTR (26). Thus, the development of experimental approaches based on Sp1 decoys to modulate the transcription of Sp1-dependent genes appears to be of great interest (3).Rec...

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Section: Synthetic Oligonucleotides and Peptidementioning

confidence: 99%

Transcription Factor Decoy Molecules Based on a Peptide Nucleic Acid (PNA)-DNA Chimera Mimicking Sp1 Binding Sites

Borgatti¹,

Lampronti²,

Romanelli³

et al. 2003

Journal of Biological Chemistry

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“…18 In order to determine the ability of the tested compounds to inhibit the cell growth and induce the erythroid differentiation, K562 cells (30,000 cells/mL) were cultured both in the absence and in the presence of the indicated concentrations of compounds and the cell number/mL determined with a ZF Coulter Counter (Counter Electronics, Hialeah, FL, USA) at different days from the culture set-up. In order to verify possible effects on erythroid differentiation, the proportion of benzidine-positive K562 cells was determined and compared to the values obtained employing other known inducers of erythroid differentiation, including cytosine arabinoside (ara-C), 19 mithramycin, 3 rapamycin 3 and butyric acid.…”

Section: Biological Activitymentioning

confidence: 99%

Synthesis of glycose carbamides and evaluation of the induction of erythroid differentiation of human erythroleukemic K562 cells

Landi

Catelani

D’Andrea

et al. 2009

European Journal of Medicinal Chemistry

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A series of carbamides derived from 1,2:5,6-di-O-isopropylidene-D-gluco-(1) and D-allofuranose (3) as well as their 5,6-O-deprotected analogues (2 and 4) and methyl 3,4-O-isopropylidene-α-and β-D-galactopyranosides (5 and 6) have been prepared in order to evaluate their ability to induce erythroid differentiation of human erythroleukemic K562 cells. Twenty out of the 51 carbamides tested exhibit an appreciable activity as inducers of erythroid differentiation and have been fully characterised and described.

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“…This cell line, which was isolated and characterized in 1975 by Lozzio et al from a patient with chronic myelogenous leukemia in blast crisis, exhibits a low proportion of Hb-synthesizing cells under standard culture conditions, but is capable of undergoing erythroid differentiation when treated with a variety of compounds including hemin (30), cytosine arabinoside (ara-C) (12), 5-azacytidine (20), chromomycin and mithramycin (12), tallimustine (10,14), cisplatin and cisplatin analogs (13). Following the erythroid induction of K562 cells, Hb Portland (˙2Á 2 ) and Hb Gower 1 (˙2ε 2 ) accumulate, due to an increase in the expression of human ˙-, ε-and Á-globin genes (20). In vitro studies demonstrated that known inducers of erythroid differentiation in K562 cells such as HU, butyrates and 5-azacytidine, are also capable of inducing HbF production when administered individually or in combination with normal erythroid cells (1).…”

Section: Discussionmentioning

confidence: 99%

“…K562 cells and cell culture conditions. The human chronic myelogenous K562 cell line (19) was obtained from the American Type Culture Collection (ATCC, Rockville, MD) and grown in vitro in RPMI-1640 medium (Sigma/Aldrich), supplemented with 10% fetal bovine serum (FBS) (Celbio, Milan, Italy), 2 mM L-glutamine (Sigma/Aldrich), a solution of 50 U/ml penicillin and 50 μg/ml streptomycin (Sigma/ Aldrich) in a humidified atmosphere of 5% CO 2 /air at 37˚C (20). K562 cells were usually seeded at 30,000 cells/ml and then treatment with bis-epoxyethyl-distamycin A derivatives was carried out by adding the appropriate drug concentrations at the beginning of the cultures.…”

Section: Methodsmentioning

confidence: 99%

“…We initially analysed erythroid differentiation by benzidine/H 2 O 2 assay and Northern blotting using the human leukemia K562 cell line as a cellular model (19,20 (21,22). This methodology can be used to identify compounds stimulating HbF accumulation in erythroid progenitors obtained from normal donors as well as subjects with hematological disorders (9).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Bis-epoxyethyl derivatives of distamycin A modified on the amidino moiety: Induction of production of fetal hemoglobin in human erythroid precursor cells

Bianchi

Borgatti²,

Fibach³

et al. 1998

Int J Mol Med

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Abstract.Derivatives of distamycin A modified at the Cterminal amidine moiety and tethered to bis-epoxyethyl moieties at the N-terminal position were tested for their ability to induce erythroid differentiation in the human erythroleukemic cell line K562. None of the compounds without bis-epoxyethyl moiety were active. A comparison of the biological activity of diepoxy compounds containing different non-basic amidine-modified moieties, showed low activity of amidoxime, carbamoyl and N-methyl carbamoyl derivatives as differentiation agents. In contrast, a cyanamidine derivative, compound 3, was able to induce erythroid differentiation of K562 cells. In addition, the cyanamidine derivative 3 was able to induce HbF accumulation following treatment of cultures of erythroid precursor cells isolated from the peripheral blood of normal subjects. IntroductionThe identification and characterization of potential therapeutic agents for the treatment of ß-thalassemia and sickle cell anemia is based on the pharmacologically mediated regulation of the expression of human Á-globin genes (1-8). It is well established that an increased production of fetal hemoglobin (HbF, · 2 Á 2 ) leads to a significant improvement of the clinical status of the patients (4,5,7). Therefore, many published experiments were designed to find hormones, cytotoxic agents, hemopoietic cytokines and short fatty acids as agents able to increase HbF levels in humans (1-3,9-11).In this respect, DNA-binding drugs are very interesting, since we reported that mithramycin, cis-platinum and tallimustine analogs are potent inducers of erythroid differentiation (12-14). The same compounds have been demonstrated to increase HbF production in human erythroid precursor cells obtained from peripheral blood of normal subjects and thalassemic patients (9). In particular, we found that the distamycin analog tallimustine and some related compounds are powerful inducers of differentiation and Á-globin mRNA accumulation in normal human erythroid progenitors (10).In this study, we report on the biological characterization of potential HbF inducers of the hybrid molecules 3-6 that include the amidine-modified tripeptide tris-N-methyl-pyrrolecarboxamide distamycin A, which contains two electrophile epoxydic functions on the amino end. In the bis-epoxyethyldistamycin A hybrids, the amidino moiety has been replaced by various non-basic groups of a different electronic nature, lipophilicity and bulk, such as cyanamidine (compound 3), amidoxime (compound 4), carbamoyl (compound 5) and Nmethyl carbamoyl (compound 6).As was previously reported, the synthesis of amidinomodified analogs provides a number of advantages (15-18). First, they are not hygroscopic and are easy to handle. Moreover, these derivatives are uncharged. Thus, products and intermediates can be readily purified by column chromatography and recrystallized.These compounds were investigated for potential erythrodifferentiation ability, in order to verify whether some of them were able to increase Á-globin gene express...

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Human leukemia K-562 cells: induction of erythroid differentiation by 5-azacytidine

Cited by 64 publications

References 24 publications

Transcription Factor Decoy Molecules Based on a Peptide Nucleic Acid (PNA)-DNA Chimera Mimicking Sp1 Binding Sites

Transcription Factor Decoy Molecules Based on a Peptide Nucleic Acid (PNA)-DNA Chimera Mimicking Sp1 Binding Sites

Synthesis of glycose carbamides and evaluation of the induction of erythroid differentiation of human erythroleukemic K562 cells

Bis-epoxyethyl derivatives of distamycin A modified on the amidino moiety: Induction of production of fetal hemoglobin in human erythroid precursor cells

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