title: Leukemia cells are sensitized to temozolomide, carmustine and melphalan by the inhibition of O6-methylguanine-DNA methyltransferase.
abstract: The cytotoxicity of the monofunctional alkylator, temozolomide (TMZ), is known to be mediated by mismatch repair (MMR) triggered by O6-alkylguanine. By contrast, the cytotoxicity of bifunctional alkylators, including carmustine (BCNU) and melphalan (MEL), depends on interstrand crosslinks formed through O6-alkylguanine, which is repaired by nucleotide excision repair and recombination. O6-alkylguanine is removed by O6-methylguanine-DNA methyltransferase (MGMT). The aim of the present study was to evaluate the cytotoxicity of TMZ, BCNU and MEL in two different leukemic cell lines (HL-60 and MOLT-4) in the context of DNA repair. The transcript levels of MGMT, ERCC1, hMLH1 and hMSH2 were determined using reverse transcription-quantitative polymerase chain reaction. In addition, the proliferation was measured using the trypan blue exclusion assay. Drug sensitivity was found to vary between the two cell lines. Treatment of the cells with TMZ, BCNU or MEL in combination with O6-benzylguanine, an MGMT inhibitor, was demonstrated to sensitize the two cell lines to these agents. However, the extent of sensitization was not found to be correlated with the expression levels of MGMT transcripts. Furthermore, the drug sensitivity was also not associated with the transcript levels of ERCC1, hMLH1 and hMSH2. Thus, leukemic cells were sensitized to alkylating agents by the inhibition of MGMT.
authors.affiliation: Department of Hematology and Oncology, Faculty of Medical Sciences, University of Fukui, Eiheiji, Fukui 910-1193, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Arai
authors.affiliation: Department of Hematology and Oncology, Faculty of Medical Sciences, University of Fukui, Eiheiji, Fukui 910-1193, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yamauchi
authors.affiliation: Department of Hematology and Oncology, Faculty of Medical Sciences, University of Fukui, Eiheiji, Fukui 910-1193, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Uzui
authors.affiliation: Department of Hematology and Oncology, Faculty of Medical Sciences, University of Fukui, Eiheiji, Fukui 910-1193, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Ueda
keywords: DNA repair; O6-benzylguanine; carmustine; melphalan; temozolomide