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Ultrastructural morphology and localisation of cisplatin-induced platinum-DNA adducts in a cisplatin-sensitive and -resistant human small cell lung cancer cell line using electron microscopy

Meijer, C., van Luyn, MJA., Nienhuis, EF., Blom, N., Mulder, NH. & de Vries, EGE., 1-Mar-2001, In : Biochemical Pharmacology. 61, 5, p. 573-578 6 p.

Research output: Contribution to journalArticleAcademicpeer-review

Ultrastructural morphology (transmission electron microscopy) and localisation of cisplatin-induced platinum (Pt)-DNA adducts (immunoelectron microscopy) were analysed in the human small cell lung cancer cell line GLC(4) and its 40-fold in vitro acquired cisplatin-resistant subline GLC(4)-CDDP, which is characterised by among other things, a decreased DNA platination. Immunolabelling of Pt-DNA adducts was performed with the polyclonal antibody GPt,known to detect the main Pt-containing intrastrand and interstrand DNA adducts, Morphological analysis of GLC(4) and GLC(4)-CDDP at the ultrastructural level showed cells with a high nucleus/cytoplasm ratio with the majority of nuclei containing one or more nucleoli. GLC(4)-CDDP showed, in contrast to GLC(4), an extensive Golgi apparatus and an increased number of mitochondria DNA platination was detectable in both GLC(4) and GLC(4)-CDDP. Immunoelectron microscopy showed Pt-DNA adducts primarily in the nucleus, preferentially at loci with high-density chromatin (e.g. heterochromatin, pars granulosa around nucleoli, condensed DNA in proliferating and apoptotic cells), and in mitochondria. The level of detectable Pt-DNA adducts was cell cycle status-dependent. In both cell lines, Pt-DNA adduct levels increased from non-dividing interphase cells to dividing cells and were highest in cells undergoing apoptosis. Overall localisation of Pt-DNA adducts was comparable in GLC(4) and GLC(4)-CDDP cells. (C) 2001 Elsevier Science Inc. All rights reserved.

Original languageEnglish
Pages (from-to)573-578
Number of pages6
JournalBiochemical Pharmacology
Volume61
Issue number5
Publication statusPublished - 1-Mar-2001

    Keywords

  • ultrastructural morphology, electron microscopy, ultrastructural localisation, cisplatin, Pt-DNA adducts, immunoelectron microscopy, CHINESE-HAMSTER OVARY, MITOCHONDRIAL-DNA, PHOTODYNAMIC THERAPY, PREFERENTIAL BINDING, ACQUIRED-RESISTANCE, CIS-DIAMMINEDICHLOROPLATINUM(II), MECHANISMS

ID: 3927953