Die Nutzung hepatischer Funktionen fuer In-vitro-Verfahren zur Pruefung von Stoffen mit dem Ziel der Einsparung von Versuchstieren. T. 8: Gentechnische Konstruktion und Nutzung von Zellinien, die hepatische Sulfotransferasen des Menschen und von Versuchstieren stabil exprimieren Schlussbericht

cDNAs encoding rat and human xenobiotic-metabolizing sulfotransferases were cloned and stably expressed in Chinese hamster V79 cells, alone or in combination with a cDNA of a cytochrome P450. The heterologously expressed enzymes showed the same electrophoretic properties and immunoreactivities as the naturally expressed enzymes from rat and human tissues. Enzyme activities towards various substrates were determined in intact cells as well as their subscellular preparations using radiometric and mass spectrometric methods. Activities were compared between cells grown under different culture conditions (passages after establishment of the recombinant cell line, presence of selecting agents, cell density, serum batch). Reference values for enzyme activities were established. Furthermore, the cells were used for the detection of sulfotransferase-mediated cyototoxic effects and genotoxic effects (gene mutations, sister chromatid exchange, DNA adducts) of chemicals. The systems were established and optimized using various pro-genotoxicants, such as 2-acetylaminofluorene and its N-hydroxylated derivative, #alpha#-hydroxytamoxifen, primary and secondary benzylic alcohols, and 2-nitropropane. (orig.)Available from TIB Hannover: F99B43 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekSIGLEBundesministerium fuer Bildung, Wissenschaft, Forschung und Technologie, Bonn (Germany)DEGerman