30 KB78 KB27 KB200520252005Plaper, Andrejaštevilka:25 straniletnik:56str. 115-119ISSN:0014-8229COBISSID:1800049URN:URN:NBN:SI:doc-KZAU7KVAslSlovensko farmacevtsko društvoFarmacevtski vestniklimfommutagenostDoločanje mutagenega potenciala snovi na celični liniji mišjega limfoma|In everyday life we are exposed to many chemicals which express different toxic potential to our organism. One of them is genotoxicity. In this paper, one of the methods which help to determine the genotoxic potential of chemicals is described. The method is mutagenicity assay on mouse lymphoma cell line (L5178Y tk+ž- 3.7.2C), heterozygous at the thymidine kinase locus (Tk1) on chromosome 11. Mutation on this part of the chromosome and inactivation of the tk+ allele induces trifluorothymidine (TFT) resistance, and tk-ž- mutants can be selected in a background of tk+ž- non-mutant cells. Mutant cells, grown in the media containing the selective substance TFT, are quantified after a certain period of time, during which the cells express the new phenotypic characteristic. The purpose and aim of the test method is a demonstration of the test material potential to induce forward mutations at the tk+tk- locus. The method is one of the mutagenicity tests included in the standard test battery for genotoxicity by regulatory agencies, and is obligatory for marketing authorisation of pharmaceuticalsV vsakdanjem življenju smo izpostavljeni številnim kemikalijam, ki na nas organizem delujejo z različno mero škodljivosti. Ena od toksičnih potencialov kemikalij je genotoksičnost. V prispevku je opisana ena od metod,s pomočjo katere prikažemo in ovrednotimo genetske poškodbe na sesalski celični liniji mišjega limfoma L5178Y tk+/- (klon 3.7.2C). Celična linija je heterozigotna na Tk1 lokusu timidinske kinaze kromosoma 11. Mutacija v tem predelu kromosoma in inaktivacija tk + alela povzroči izgubo aktivnosti tega encima in s tem pridobitev rezistence na tri-fluorotimidin (TFT). Tako lahko mutante tk-/- ločimo od nemutiranih celic. Mutirane celice,ki rastejo v selektivnem gojišču s TFT, kvantificiramo po določenem času, v katerem izrazijo svojo novo fenotipsko lastnost. Namen in cilj metode je prikazati potencial testne substance, da inducira mutacije na lokusu tk+tk-. Metoda je ena od testov mutagenosti, ki jih regulatorne organizacije uvrščajo v standardno skupino testov za določanje genotoksioenega potenciala zdravilnih učinkovin, ki morajo biti izvedeni pred registracijoTEXTznanstveno časopisjejournalsSlovenian National E-content AggregatorNational and University Library of SloveniaSlovensko farmacevtsko društvo