Assessment of genistein influence on TP53 and CDKN1A expression in keratinocytes cultured as monolayer and multilayer culture exposed to UV lightKatarzyna Adamczyk 1/, Magdalena Jurzak 1/, Małgorzata Latocha 2/, Dariusz Kuśmierz 2/, Paweł Antończak 1/, Agnieszka Garncarczyk 1/ 1/ Zakład Kosmetologii Katedry Kosmetologii, Wydział Farmaceutyczny z Oddziałem Medycyny Laboratoryjnej w Sosnowcu, Śląski Uniwersytet Medyczny w Katowicach 2/ Zakład Biologii Komórki, Wydział Farmaceutyczny z Oddziałem Medycyny Laboratoryjnej w Sosnowcu, Śląski Uniwersytet Medyczny w Katowicach Summary Introduction. UV radiation is one of major factors accelerating skin aging and inducing process of carcinogenesis. Preventing adverse effects of exposure to UV radiation requires the use of photoprotecting cosmetics. The search for new active ingredients requires research using in vitro cultures of skin cells. Aim. Aim of this study was to evaluate the effect of genistein on the expression of TP53 and CDKN1A genes in epidermal keratinocytes in monolayer cultures and epidermis equivalents that were exposed to UV radiation. Material and methods. Material consisted human epidermal keratinocytes and epidermis equivalent EpiDerm™. Cells viability treated with genistein was assayed with the use of WST-1. The copy number of TP53 and CDKN1A mRNA in extracts from keratinocytes and epidermal equivalents incubated 72-hours with genistein at 1.85 μM and 18.5 μM concentrations and 15 minutes long UV irradiated was determined by performing quantitative RT-PCR reactions using QuantiTech™SYBR®Green™ RT-PCR kit. Results. After exposure to UV radiation, in both types of culture increased expression of TP53 gene was observed. However only in keratinocytes an increase in expression of CDKN1A was observed. Incubation with genistein before exposure to UV radiation differently modulate the expression of the genes in keratinocytes and epidermis equivalents. The results did not allow for a clear assessment of the photoprotecting potential of genistein. Conclusion. The search for new photoprotecting substances requires research on a variety of biological material. In order to determine the photoprotecting potential of genistein it is necessary to conduct further studies on the effect of the flavonoid on the expression of UV-response genes in more trials. More genes should be examined us well. Key words: genistein, TP53, CDKN1A, keratinocytes, epidermis model EpiDerm™, UV light |