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Mutagenesis, Vol. 17, No. 4, 317-320, July 2002
© 2002 UK Environmental Mutagen Society/Oxford University Press

Cell studies of the DNA bis-intercalator {Delta}-{Delta} [µ-C4(cpdppz)2-(phen)4Ru2]4+: toxic effects and properties as a light emitting DNA probe in V79 Chinese hamster cells

Björn Önfelt2, Lovisa Göstring1, Per Lincoln, Bengt Nordén and Agneta Önfelt1

Department of Physical Chemistry, Chalmers University of Technology, S-412 96 Gothenburg, Sweden and 1 Genetic and Cellular Toxicology, Wallenberg Laboratory, Stockholm University, S-106 91 Stockholm, Sweden

Coordination complexes of type [Ru(L)3]2+, where L is a nitrogen-containing aromatic bidentate ligand, can often be photolytically reduced, making them useful in studies of DNA- or protein-mediated electron transfer and in artifical photosynthesis model systems. Upon binding to DNA some Ru(L) complexes have been found to display strongly increased fluorescence compared with when free in solution, making those compounds interesting to test as DNA probes. Thus, they are becoming widely used in the chemistry community. Here, asynchronous cultures of V79 Chinese hamster cells were exposed to the DNA bis-intercalator {Delta}{Delta}-{Delta}{Delta} [µ-C4(cpdppz)2-(phen)4Ru2]4+ at 10-10–10-4 M. The extraordinarily strong binding of the compound to DNA was the reason for testing its possible interference with DNA metabolism in intact mammalian cells. Exposure for 1 h to 10-10–10-4 M did not significantly decrease DNA synthesis. Cells exposed to 10-5 M for 27 h showed no staining of the nucleus, while DNA was stained in cells electroporated in the presence of the compound. However, the Ru dimer was probably taken up by pinocytosis, because numerous minute precipitates could be observed in the cytoplasm. Treatment for 24 h at concentrations of 10-10–10-5 M did not inhibit growth, as indicated by cell density and mitotic activity. Neither did it affect chromosomal arrangements during mitosis. However, at 10-4 M the density of cultures was reduced by ~45% and apoptotic cells were frequent, as opposed to mitoses. We also investigated the properties of the Ru dimer as a fluorescent DNA stain. The compound appears attractive as a red DNA stain when broad excitation in the visible range is desirable and extremely low background staining is essential. The low toxicity of the compound is a favourable trait in this context.

2 To whom correspondence should be addressed. Tel: +46 31 7725120; Fax: +46 31 7723858; Email: onfelt{at}phc.chalmers.se


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