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

  • A. Freund
  • A. Gescher
  • J. Boos
Conference paper
Part of the Haematology and Blood Transfusion / Hämatologie und Bluttransfusion book series (HAEMATOLOGY, volume 39)

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.

Keywords

Acute Myeloid Leukemia HL60 Cell Cytosine Arabinoside Human Myeloid Leukemia Cell Relapse Acute Myeloid Leukemia 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • A. Freund
    • 1
  • A. Gescher
    • 2
  • J. Boos
    • 1
  1. 1.Dept. of Pediatric OncologyUniversity of MünsterGermany
  2. 2.Medical Research Council Toxicology UnitUniversity of LeicesterUK

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