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Modulation of Ara-C Cytotoxicity by Coadministration with Antisignalling Drugs in HL60 and Ara-C-Resistant HL60/Ara-C Cells

  • Conference paper
Acute Leukemias VII

Part of the book series: Haematology and Blood Transfusion / Hämatologie und Bluttransfusion ((HAEMATOLOGY,volume 39))

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Abstract

Ara-C (cytosine arabinoside) is one of the most effective drugs in the treatment of acute myeloid leukemia (AML). Ara-C is phosphorylated to Ara-CTP (cytosine arabinoside triphosphate) which inhibits DNA polymerase and induces DNA strand breaks after incorporation into DNA. There is increasing evidence that ara-C also affects some cellular signal transduction pathways; it induces transcription/expression of c-fos, c-jun, NF kappaB, protein kinase C δ, mitogen-activated protein kinase and cyclin E and downregulates cyclin-dependent-kinase-2 and retinoblastoma protein phosphorylation. These events may contribute to ara-C cytotoxicity. The antisignalling drugs all-trans retinoic acid (ATRA), staurosporine, quercetin and bryostatin-1 have recently been shown to enhance ara-C cytotoxicity.

We chose 7 different antisignalling agents [ATRA and the kinase inhibitors quercetin, genistein, CGP 52411, tyrphostin A48, nordihydroguaiaretic acid (NDGA) and staurosporine] and compared their ara-C sensitising potencies with those of hydroxyurea (HU), arabinosyl-2-fluoroadenine (F-Ara-A) and 2-chlorodeoxyadenosine (2-CdA), agents currently in use as “second line treatment” in resistant and relapsed AML. Cytotoxicity was assessed by the tetrazolium (MTT) assay in HL60 cells and a newly derived ara-C-resistant subline (HL60/ara-C) in at least three separate experiments. Supraadditive cytotoxicity was found for combinations containing ATRA, CGP 52411, tyrphostin A48 and HU in both cell lines, for 2-CdA, staurosporine and NDGA in HL60, and for F-Ara-A in HL60/ara-C cells. Quercetin and genistein did not sensitise cells against ara-C.

To elucidate the mechanism of sensitisation in HL60 cells we studied the influence of the modulators of ara-C cytotoxicity on cellular markers of apoptosis. After 4h-coincubation we measured cell size (volume), DNA loss by flow cytometry (sub-G1 peak) and DNA fragmentation by conventional gel electrophoresis. Tyrphostin A48, NDGA, ATRA and HU increased ara-C-induced apoptosis, whereas staurosporine did not affect it. CGP 52411 decreased the effect of ara-C on apoptotic indicators after 4h-, but no longer after 12h-coincubation.

The results suggest that antisignalling drugs such as ATRA, CGP 52411, tyrphostin A48, staurosporine and NDGA may be efficacious alternatives to the already clinically applied ara-C modulators. Among the clinically used ara-C modulators, HU sensitised more potently against ara-C than F-Ara-A and 2-CdA.

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Authors and Affiliations

  1. Dept. of Pediatric Oncology, University of Münster, Germany

    A. Freund & J. Boos

  2. Medical Research Council Toxicology Unit, University of Leicester, UK

    A. Gescher

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  1. A. Freund
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  2. A. Gescher
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  3. J. Boos
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Editor information

Editors and Affiliations

  1. Department of Internal Medicine, University of Göttingen, Robert-Koch-Straße 40, 37075, Göttingen, Germany

    W. Hiddemann  & B. Wörmann  & 

  2. Department of Internal Medicine, USA

    T. Büchner

  3. Department of Pediatrics, University of Münster, Albert-Schweitzer-Straße 33, 48149, Münster, Germany

    J. Ritter  & U. Creutzig  & 

  4. Dept. of Hematology, University of Texas, Anderson Cancer Center, 1515 Holcombe Boulevard, 77030, Houston, Tx., USA

    M. Keating MD PhD  & W. Plunkett MD PhD  & 

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© 1998 Springer-Verlag Berlin Heidelberg

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Freund, A., Gescher, A., Boos, J. (1998). Modulation of Ara-C Cytotoxicity by Coadministration with Antisignalling Drugs in HL60 and Ara-C-Resistant HL60/Ara-C Cells. In: Hiddemann, W., et al. Acute Leukemias VII. Haematology and Blood Transfusion / Hämatologie und Bluttransfusion, vol 39. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-71960-8_83

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  • DOI: https://doi.org/10.1007/978-3-642-71960-8_83

  • Publisher Name: Springer, Berlin, Heidelberg

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