Analysis of blastomogenic activity of mammal carcinogens in Drosophila using the wts ~(P4) allele and RNA interference-induced P53 silencing

摘要

The test for somatic mutagenesis and recombination in Drosophila is one of the widely used approaches for determination of possible carcinogenic effects of chemical compounds. The use of heterozygotes for mutant tumor suppressor gene wts enables more direct evaluation of the blastomogenic effects of chemical compounds, by tumor formation in the adult flies. This study presents evaluation of the SMART effectiveness upon the use of Drosophila heterozygotes for the wts~(P4) gene, first included into the test system. The increase of the test resolution capacity compared to the literature data for the wts~(P2) allele was observed. Using wts~(P4) heterozygotes, a total of 20 carcinogenic compounds, and their slightly carcinogenic and non-carcinogenic analogs were tested. Specificity of the method was about 100%, and sensitivity depended on the type of the agent tested. The latter was absolute for the direct action carcinogens, with respect to carcinogens, requiring the metabolic activation. The sensitivity was elective and was limited by the presence of the enzymes capable of activating of these compounds. To increase the test sensitivity, the RNA interference-mediated silencing of the Drosophila p53 functional activity was successfully applied. Moreover, the frequency of wts tumor induction considerably increased both in spontaneous and induced mutagenesis conditions.