Background. The mechanism of cisplatin-induced nephrotoxicity is not well understood. The distal tubules are affected in both human and animal studies, although the majority of cisplatin-induced renal damage is in proximal tubules. Platinum (Pt) forms intra- and interstrand cross-links with DNA in cancer cells. Hypothesis. A mechanism of cisplatin-induced cytotoxicity in MDCK cells relates to its ability to bind to DNA and interfere with its synthesis. Methods. The canine distal renal tubular epithelial cell line, MDCK was used as an in vitro model to investigate the mechanism of cisplatin-induced nephrotoxicity. The intracellular Pt accumulation and Pt binding with DNA were assayed by atomic absorption spectrophotometry. DNA synthesis was measured by BrdU labeling and fluorescence microscopy at the concentrations from 0 to 100 muM. The alkaline comet assay with 10 Gy radiation was used to measure Pt DNA interstrand cross-links after a one hour cisplatin exposure from 0 to 100 muM at both zero and sixteen hour time points. According to the principles of alkaline comet assay, the tail moment is inversely related with the amount of Pt interstrand cross-links. (Abstract shortened by UMI.)
| Identifer | oai:union.ndltd.org:uottawa.ca/oai:ruor.uottawa.ca:10393/6408 |
| Date | January 2002 |
| Creators | Li, Yan Julia. |
| Contributors | Goel, Rakesh, |
| Publisher | University of Ottawa (Canada) |
| Source Sets | Université d’Ottawa |
| Detected Language | English |
| Type | Thesis |
| Format | 88 p. |
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