Induction of differentiation in the human promyelocytic leukemia cell line HL-60 by the cyclopentenyl analogue of cytidine

Glazer, Robert I.; Cohen, Marvin B.; Hartman, Kathleen D.; Knode, Marian C.; Lim, Mu-Ill; Márquez, Víctor E.

doi:10.1016/0006-2952(86)90301-1

Cited by 39 publications

(10 citation statements)

References 17 publications

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“…CTP is the immediate precursor of the activated, energy-rich phospholipid pathway intermediates CDP-diacylglycerol, CDP-ethanolamine, and CDP-choline, which are essential intermediates in phospholipid synthesis during cell cycle progression (Jackowski et al, 2000). Treatment of promyelocytic HL-60 cells with an inhibitor of CTP synthase (e.g., cyclopentenyl cytosine) (Ford et al, 1991), induced both growth inhibition and differentiation of these cells that was accompanied by a pronounced decline in the level of CTP, but not of UTP, ATP, or GTP (Glazer et al, 1986). In related studies, we have observed that cyclopentenyl cytosine also induces a dose-dependent differentiation of K562 cells (M. Huang, Y. H. Wang, M. Collins, and L. M. Graves, unpublished observations).…”

Section: Discussionmentioning

confidence: 99%

A77 1726 Induces Differentiation of Human Myeloid Leukemia K562 Cells by Depletion of Intracellular CTP Pools

et al. 2002

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A77 1726 (LEF) is the active metabolite of leflunomide, a recently approved immunosuppressive agent. We examined the ability of LEF to induce differentiation of a human erythroleukemia (K562) cell line and show that LEF induces a dose-and time-dependent differentiation of these cells as characterized by growth inhibition, hemoglobin production, and erythroid membrane protein glycophorin A expression. This effect was dependent on depletion of the intracellular pyrimidine ribonucleotides (UTP and CTP), and preceded by a specific Sphase arrest of the cell cycle. Supplementation of the cultures with exogenous uridine restored intracellular UTP and CTP to normal levels and prevented the LEF-induced cell cycle block and differentiation of K562 cells. Interestingly, addition of cytidine alone blocked the LEF-induced differentiation of K562 cells but only restored the CTP pool. By contrast, neither deoxycytidine nor thymidine prevented the effects of LEF on these cells. Similarly, pyrimidine starvation of a cell line lacking the de novo pyrimidine pathway (G9c) resulted in an S-phase arrest that was reversed by the addition of cytidine. Thus these studies demonstrate an important role for CTP in regulating cell cycle progression and show that LEF is an effective inducer of tumor cell differentiation through depletion of this ribonucleotide.

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Section: Discussionmentioning

confidence: 99%

A77 1726 Induces Differentiation of Human Myeloid Leukemia K562 Cells by Depletion of Intracellular CTP Pools

et al. 2002

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“…17 Cyclopentenyl cytosine (CPEC) is a nucleoside analogue that inhibits CTP synthetase, inducing a depletion of CTP and dCTP 18 -23 in several cell lines of solid tumors and hematologic malignancies, resulting in a decreased synthesis of DNA and RNA. 19,22 In mice, CPEC proved to have an anti-tumor effect in vivo. 18,24,25 Being a cytidine analogue, CPEC is phosphorylated thrice into its active metabolite CPEC-triphosphate (-TP) that inhibits CTP synthetase.…”

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confidence: 99%

Cyclopentenyl cytosine increases the phosphorylation and incorporation into DNA of 1‐β‐D‐arabinofuranosyl cytosine in a human T‐lymphoblastic cell line

Verschuur

Gennip

Leen

et al. 2002

Intl Journal of Cancer

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The cytotoxic effect of 1-␤-D-arabinofuranosyl cytosine (araC) depends on the intracellular phosphorylation into its active compound araCTP, on the degree of degradation of araCTP and on the incorporation of araCTP into DNA. Deoxycytidine triphosphate (dCTP) inhibits the phosphorylation of araC (by feedback inhibition of the enzyme deoxycytidine kinase) and the incorporation of araCTP into DNA (by competition for DNA polymerase). In a T-lymphoblastic cell line, we studied whether the cytotoxicity of araC (2 nM-50 M) could be enhanced by decreasing the concentration of dCTP, using the nucleoside-analogue cyclopentenyl cytosine (CPEC), an inhibitor of the enzyme CTP synthetase. Preincubation of the cells with CPEC (100 -1,600 nM) for 2 hr increased the concentrations of araCMP 1.6 -9.5-fold, which was significant for each concentration of CPEC used. The concentration of araCDP remained low, whereas the concentration of araCTP changed depending on the concentration of araC used. With 2-15 M of araC and a preincubation with 400 nM of CPEC, the araCTP concentration increased by 4 -15% (not significant), and the total amount of araC nucleotides increased significantly by 21-45%. When using a concentration of araC of 2 nM after a preincubation with CPEC of 100 nM, the concentration of araCMP increased by 60% (p ‫؍‬ 0.015), whereas that of araCTP decreased by 10% (p ‫؍‬ 0.008). This was compensated by an increase of 41% (p ‫؍‬ 0.005) of araCTP incorporation into DNA, which represented 43% of all araC metabolites. Moreover, by performing pulse/chase experiments with 400 nM of CPEC and 2 M of araC, the retention of cytosolic araCTP and the incorporated amount of araCTP into DNA were increased by CPEC. The modulation by CPEC of araC metabolism was accompanied by a synergistic increase of araC-induced apoptosis and by an additive effect on the araC-induced growth inhibition. © 2002 Wiley-Liss, Inc. Key words: cytarabine; deoxycytidine kinase; dCTP; CTP synthetase; cyclopentenyl cytosine 1-␤-D arabinofuranosyl cytosine (araC) is a nucleoside analogue that constitutes the main drug of virtually all treatment protocols for AML and proved its benefit in precursor-B acute lymphocytic leukemia (ALL) and T-ALL. 1-3 One can distinguish low-dose (LD-araC) regimens, applying doses of 75-200 mg/m 2 , from high-dose (HD-araC) regimens where 1-3 g/m 2 are used. The cytotoxic effect of araC depends on several intracellular mechanisms: araC is metabolized into arabinofuranosyl cytosine triphosphate (araCTP) by 3 consecutive phosphorylating steps, catalyzed by deoxycytidine kinase (dCK), nucleoside monophosphate (NMP) kinase and nucleoside diphosphate (NDP) kinase, respectively (Fig. 1). AraCTP is incorporated into DNA as a deoxynucleotide-analogue, where it exerts its cytotoxic properties by inhibiting DNA replication. 4 -6 The phosphorylation of araC into araCTP can be counteracted by the enzyme (deoxy)cytidine deaminase, which converts araC into arabinofuranosyl uracil (araU), dCMP deaminase, converting arabinofuranosyl cytosine mon...

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“…The depletion of CTP and deoxy-CTP is expected to be the mechanism through which CPEC reduces the synthesis of nucleic acids (Glazer et al, 1986). In addition, the salvage pathway of uridine was inhibited by CPEC, which may also contribute to the decreased RNA synthesis and the cytostatic effect induced by CPEC (Glazer et al, 1986;Ford et al, 1991). In terms of CTP depletion, a dose±response effect was obtained with CPEC.…”

Section: Discussionmentioning

confidence: 99%

“…In a previous study, a high CTP synthetase activity was demonstrated in lymphoblasts of a limited number of paediatric patients with ALL (Verschuur et al, 1998). For these reasons, CTP synthetase might be an attractive target for chemotherapy because inhibition of the enzyme leads to a CTP and dCTP depletion, resulting in a reduced synthesis of nucleic acids (Glazer et al, 1986;Ford et al, 1991). Furthermore, inhibition of CTP synthetase may increase the cytotoxicity of some nucleoside analogues, such as arabinofuranosyl cytosine (Grem & Allegra, 1990, Grem et al, 1991.…”

mentioning

confidence: 99%

“…It has been shown to reduce DNA and RNA synthesis in lymphocytic (Ford et al, 1991) and myeloid (Glazer et al, 1986) leukaemic cell lines. In addition, a growth-inhibiting effect of CPEC has been observed on several malignant cell lines, including a human (De Clerq et al, 1991;Ford et al, 1991) and a murine lymphocytic leukaemic cell line (Kang et al, 1989;Ford et al, 1995).…”

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confidence: 99%

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In vitro inhibition of cytidine triphosphate synthetase activity by cyclopentenyl cytosine in paediatric acute lymphocytic leukaemia

Verschuur

Gennip²,

Leen³

et al. 2000

Br J Haematol

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Summary. Cytidine triphosphate (CTP) synthetase is a key enzyme for the synthesis of cytosine (deoxy)ribonucleotides, catalysing the conversion of uridine triphosphate (UTP) into CTP, and has a high activity in several malignancies. In this preclinical study, the enzyme activity and mRNA expression of the enzyme and (deoxy)ribonucleotide concentrations were analysed in leukaemic cells of 57 children suffering from acute lymphocytic leukaemia (ALL). In addition, in vitro experiments were performed with the CTP synthetase inhibitor cyclopentenyl cytosine (CPEC). A significantly higher activity of CTP synthetase (6´5^3´9 nmol CTP/ mg/h) was detected in ALL cells than in lymphocytes of healthy controls (1´8^0´9 nmol CTP/mg/h, P , 0´001) that was independent of white blood cell (WBC) count, blast percentage, age, gender or type of ALL. The enzyme activity was not correlated with the CTP synthetase mRNA expression. The activity of CTP synthetase in ALL cells compared with non-malignant CD341 bone marrow controls (5´6^2´4 nmol CTP/mg/h) was not statistically different. In vitro treatment of ALL cells with CPEC induced a dose-dependent decrease of the CTP concentration. The lowest concentration of CPEC (0´63 mm) induced a depletion of CTP of 41^20% and a depletion of dCTP of 27^21%. The degree of CTP depletion of ALL cells after treatment with CPEC was positively correlated with the activity of CTP synthetase. The inhibition of CTP synthetase in situ was confirmed by flux studies using radiolabelled uridine. From these results, it can be expected that CPEC has a cytostatic effect on lymphoblasts of children with ALL.

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Induction of differentiation in the human promyelocytic leukemia cell line HL-60 by the cyclopentenyl analogue of cytidine

Cited by 39 publications

References 17 publications

A77 1726 Induces Differentiation of Human Myeloid Leukemia K562 Cells by Depletion of Intracellular CTP Pools

A77 1726 Induces Differentiation of Human Myeloid Leukemia K562 Cells by Depletion of Intracellular CTP Pools

Cyclopentenyl cytosine increases the phosphorylation and incorporation into DNA of 1‐β‐D‐arabinofuranosyl cytosine in a human T‐lymphoblastic cell line

In vitro inhibition of cytidine triphosphate synthetase activity by cyclopentenyl cytosine in paediatric acute lymphocytic leukaemia

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