title: Effects of single or split exposure of leukemic cells to temozolomide, combined with poly(ADP-ribose) polymerase inhibitors on cell growth, chromosomal aberrations and base excision repair components.
abstract: OBJECTIVE
  To evaluate the antitumor activity of single versus split exposure of neoplastic cells to temozolomide (TZM) and poly(ADP-ribose) polymerase (PARP) inhibitor.
  
  
  METHODS
  A leukemic Jurkat cell line and freshly isolated leukemic blasts were used. Jurkat cells are resistant to O6-methylguanine damage induced by TZM due to high levels of O6-alkylguanine-DNA alkyltransferase and to a functional defect in the mismatch repair system. Cells were treated with 3-aminobenzamide or with NU1025 to inhibit PARP activity. TZM was added to cell cultures immediately after PARP inhibitors. The concentrations of TZM used were 62.5 microM (corresponding to the peak plasma concentration in patients) or 125 microM. TREATMENT DESIGN: Cells were treated with 125 microM TZM plus PARP inhibitors (single exposure), or twice with 62.5 microM TZM plus PARP inhibitors with an interval of 24 h between treatments (split exposure). Tumor cell growth, clastogenicity and base excision repair gene transcripts or enzymatic activity were evaluated.
  
  
  RESULTS
  The split exposure of Jurkat cells to TZM induced more pronounced and persistent growth inhibition and comparable chromosome damage in comparison with the single exposure. In addition, PARP inhibitors potentiated the cytotoxic effects induced by repeated treatment with TZM in fresh leukemic blasts. A marked decrease in X-ray repair cross-complementing 1 transcript and methylpurine glycosylase (MPG) transcript was detected in Jurkat cells subjected to the split exposure. In this case, a significant reduction in the corresponding enzymatic activity was also observed.
  
  
  CONCLUSIONS
  Cytotoxicity induced by TZM and PARP inhibitors can be improved by a fractionated modality of drug treatment. The reduction in MPG transcript and function would presumably contribute to an increase in cell susceptibility to DNA damage induced by the methylating agent and PARP inhibitors.
authors.affiliation: Department of Neuroscience, Section of Pharmacology and Medical Oncology, University of Rome Tor Vergata, Via di Tor Vergata 135, 00133 Rome, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Tentori
authors.forename: I
authors.initials: I
authors.lastname: Portarena
authors.forename: P
authors.initials: P
authors.lastname: Vernole
authors.forename: P
authors.initials: P
authors.lastname: De Fabritiis
authors.forename: R
authors.initials: R
authors.lastname: Madaio
authors.forename: A
authors.initials: A
authors.lastname: Balduzzi
authors.forename: R
authors.initials: R
authors.lastname: Roy
authors.forename: E
authors.initials: E
authors.lastname: Bonmassar
authors.forename: G
authors.initials: G
authors.lastname: Graziani
mesh_terms: D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D015152:Blotting, Northern; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002869:Chromosome Aberrations; D004247:DNA / Q000096:biosynthesis; D004260:DNA Repair / Q000187:drug effects*; D003606:Dacarbazine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D019169:Jurkat Cells; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D000077204:Temozolomide
chemical_list: D018906:Antineoplastic Agents, Alkylating; D004791:Enzyme Inhibitors; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D003606:Dacarbazine; D004247:DNA; D000077204:Temozolomide