title: Analysis to Estimate Genetic Variations in the Idarubicin-Resistant Derivative MOLT-3.
abstract: Gene alterations are a well-established mechanism leading to drug resistance in acute leukemia cells. A full understanding of the mechanisms of drug resistance in these cells will facilitate more effective chemotherapy. In this study, we investigated the mechanism(s) of drug resistance in the human acute leukemia cell line MOLT-3 and its idarubicin-resistant derivative MOLT-3/IDR through complete mitochondrial and nuclear DNA analyses. We identified genetic differences between these two cell lines. The ND3 mutation site (p.Thr61Ile) in the mitochondrial DNA sequence was unique to MOLT-3/IDR cells. Moreover, we identified five candidate genes harboring genetic alterations, includingGALNT2, via CGH array analysis. Sequencing of theGALNT2exon revealed a G1716K mutation present within the stop codon in MOLT-3/IDR cells but absent from MOLT-3 cells. This mutation led to an additional 18 amino acids in the protein encoded byGALNT2. Using real-time PCR, we determined an expression value for this gene of 0.35. Protein structure predictions confirmed a structural change inGALNT2in MOLT-3/IDR cells that corresponded to the site of the mutation. We speculate that this mutation may be related to idarubicin resistance.
authors.affiliation: Department of Clinical Pharmacology, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. komiyama@tokai-u.jp.
authors.forename: Tomoyoshi
authors.initials: T
authors.lastname: Komiyama
authors.affiliation: Nagahama Institute of Bio-Science and Technology, Nagahama, Shiga 526-0829, Japan. aogu@whelix.info.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ogura
authors.affiliation: The National Institute of Advanced Industrial Science and Technology (AIST), Tokyo Waterfront Bio-IT Research Building 2-4-7 Aomi, Koto-ku, Tokyo 135-0064, Japan. t-hirokawa@aist.go.jp.
authors.forename: Takatsugu
authors.initials: T
authors.lastname: Hirokawa
authors.affiliation: Department of Clinical Pharmacology, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. mrprince329@yahoo.co.jp.
authors.forename: Miao
authors.initials: M
authors.lastname: Zhijing
authors.affiliation: Support Center for Medical Research and Education, Tokai University, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. kamiguti@is.icc.u-tokai.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kamiguchi
authors.affiliation: Department of Laboratory Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. sa@is.icc.u-tokai.ac.jp.
authors.forename: Satomi
authors.initials: S
authors.lastname: Asai
authors.affiliation: Department of Laboratory Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. miyachi@is.icc.u-tokai.ac.jp.
authors.forename: Hayato
authors.initials: H
authors.lastname: Miyachi
authors.affiliation: Department of Clinical Pharmacology, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. hkobayas@is.icc.u-tokai.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kobayashi
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D045744:Cell Line, Tumor; D018388:Codon, Terminator / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005091:Exons; D006801:Humans; D015255:Idarubicin / Q000494:pharmacology*; D020125:Mutation, Missense; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics; D011110:Polymorphism, Genetic; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D000903:Antibiotics, Antineoplastic; D018388:Codon, Terminator; D004272:DNA, Mitochondrial; D015255:Idarubicin; D017350:N-Acetylgalactosaminyltransferases; D000097763:Polypeptide N-acetylgalactosaminyltransferase
keywords: CGH array; GALNT2 gene; MOLT-3; gene polymorphism; genetic variations; idarubicin (IDR); leukemia; mitochondrial DNA