Cisplatin-induced apoptosis in cultures of human Schwann cells.
To investigate the sensitivity of human Schwann cells to cisplatin (cis-DDP), different approaches to estimate DNA damage were used: the comet assay, morphological evaluation of the granular condensation of nuclear chromatin and the terminal transferase-mediated dUTP nick-end-labelling (TUNEL) method. The number of micronuclei (MNi), as a sign of cisplatin-induced genotoxicity, was counted. DNA damage assessed by the comet assay was already evident after 1.5 microM cisplatin treatment at all exposure times (24, 48, and 72 h). Initial morphological changes characterised by the granular condensation of nuclear chromatin were detectable after 24 h exposure to 25 microM cis-DDP, while an increased number of apoptotic cells, determined by the TUNEL method, was noted after 48 h exposure to the same concentration. The first significant increase in the number of MNi was observed in cells treated with 75 microM cis-DDP for 24 h. We demonstrate that the comet assay is a highly sensitive method for measuring cisplatin induced DNA damage. Morphological observation revealed advanced as well as less prominent alterations in the nuclear chromatin. In contrast, the TUNEL method detected only those cells with advanced DNA fragmentation.
Jirsova
Katerina
K
Laboratory and Ocular Tissue Bank, Department of Ophthalmology, General Teaching Hospital and Charles University, Prague, Czech Republic. katerina.jirsova@vfn.cz
Mandys
Vaclav
V
Gispen
Willem Hendrik
WH
Bär
Peter Rudolf
PR
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002452:Cell Count / Q000379:methods; D002467:Cell Nucleus / Q000187:drug effects; D002478:Cells, Cultured; D002843:Chromatin / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology*; D020552:Comet Assay / Q000379:methods; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D020287:In Situ Nick-End Labeling / Q000379:methods; D012583:Schwann Cells / Q000187:drug effects* / Q000473:pathology; D013997:Time Factors
D000970:Antineoplastic Agents; D002843:Chromatin; D002945:Cisplatin