Untersuchungen zu Mechanismen der Arsenresistenz in kultivierten V79-Zellen und daraus selektierten Klonen

Studies on mechanisms of arsenic resistance in cultured V79 cells and from selected clones

von Claudia Bässler
Dissertation
Datum der mündl. Prüfung:2013-08-05
Erschienen:2013-07-30
Betreuer:Dr. Prof .Gebel
Gutachter:Prof. Dr. Ralf Dressel
Gutachter:PD Dr. G. Westphal
Zum Verlinken/Zitieren: http://dx.doi.org/10.53846/goediss-3961

 

 

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In the present work it was worked with the established Fibroblast cell line of the Chinese hamster (V79 cells) and selected arsenite resistant clones of this cell line. 5 of the Arsenite resistant clones (K4, K6, K7, K8 K9) were investigated by the established V79 line by Neutral-red-Assay on possible increased tolerance to the Cytotoxicity of Arsenite (AsIII) and Cisplatin (CDDP) by micronucleous-test and also for possible increased tolerance to the Genotoxicity from arsenite. The Arsenite resistance ED50 in the V79 clones (K4, K6, K7, K8, K9) were increased by the Double one to fivefold. In addition a cross-resistance could be ascertained to cisplatin (CDDP). The genotoxic potential of Arsenite or the formation of microcores was coined in the Arsenite resistant V79 clones in general more slightly in comparison to the untreated V79 cells (V79). The RNA from V79 cells were used for investigate the ABC-transporter Mrp1 and Mrp2 and to the examination of the mRNA expression. The cDNA fragments got by RT-PCR were cloned and sequenced. The Mrp2 mRNA expression in the fibroblastoid lung epithelium cell V79 of the hamster could not be proved in the Northern Blot. However, in 4 of the 5 arsenite resistant clones showed an increased Mrp1 mRNA expression in comparison to the not cloned controlling cells (K). Nevertheless, with a clone (K8) a low Mrp1 mRNA expression was remarkable in comparison to the other resistant clones, although with this clone in the Neutral-red-assay a relatively high tolerance could be ascertained towards Arsenite. These results speak for the fact, that the Mrp1-transporter expression could contribute to the arsenite resistance in general. Nevertheless, in case of the clone K8 the arsenite resistance could other mechanisms also be possible.
Keywords: arsenite resistance V79-cells
Schlagwörter: Arsenresistenz V79-Zellen
 
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