Dysregulation of gap junction intercellular communication (GJIC) is recognized as one of the key hallmarks for identifying non-genotoxic carcinogens (NGTxC). Currently, there is a demand for in vitro assays addressing the gap junction hallmark, which would have the potential to eventually become an integral part of an integrated approach to the testing and assessment (IATA) of NGTxC. The scrape loading-dye transfer (SL-DT) technique is a simple assay for the functional evaluation of GJIC in various in vitro cultured mammalian cells and represents an interesting candidate assay. Out of the various techniques for evaluating GJIC, the SL-DT assay has been used frequently to assess the effects of various chemicals on GJIC in toxicological and tumor promotion research. In this review, we systematically searched the existing literature to gather papers assessing GJIC using the SL-DT assay in a rat liver epithelial cell line, WB-F344, after treating with chemicals, especially environmental and food toxicants, drugs, reproductive-, cardio- and neuro-toxicants and chemical tumor promoters. We discuss findings derived from the SL-DT assay with the known knowledge about the tumor-promoting activity and carcinogenicity of the assessed chemicals to evaluate the predictive capacity of the SL-DT assay in terms of its sensitivity, specificity and accuracy for identifying carcinogens. These data represent important information with respect to the applicability of the SL-DT assay for the testing of NGTxC within the IATA framework.
- Klíčová slova
- carcinogenesis, carcinogens, gap junction intercellular communication, scrape loading-dye transfer,
- MeSH
- barvicí látky metabolismus MeSH
- biotest metody MeSH
- buněčné linie MeSH
- fluorescenční mikroskopie metody MeSH
- játra patologie MeSH
- karcinogeny MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- mezerový spoj metabolismus MeSH
- mezibuněčná komunikace účinky léků fyziologie MeSH
- testy karcinogenity metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- barvicí látky MeSH
- karcinogeny MeSH
In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures and chemical interactions. We aimed to investigate cytotoxic and genotoxic effects of complex extracts from laboratory cultures of cyanobacterial species from different orders (Cylindrospermopsis raciborskii, Aphanizomenon gracile, Microcystis aeruginosa, M. viridis, M. ichtyoblabe, Planktothrix agardhii, Limnothrix redekei) and algae (Desmodesmus quadricauda), and examine possible relationships between the observed effects and toxin and retinoic acid (RA) content in the extracts. The cytotoxic and genotoxic effects of the extracts were studied in the human hepatocellular carcinoma HepG2 cell line, using the MTT assay, and the comet and cytokinesis-block micronucleus (cytome) assays, respectively. Liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS) was used to detect toxins (microcystins (MC-LR, MC-RR, MC-YR) and cylindrospermopsin) and RAs (ATRA and 9cis-RA) in the extracts. Six out of eight extracts were cytotoxic (0.04-2 mgDM/mL), and five induced DNA strand breaks at non-cytotoxic concentrations (0.2-2 mgDM/mL). The extracts with genotoxic activity also had the highest content of RAs and there was a linear association between RA content and genotoxicity, indicating their possible involvement; however further research is needed to identify and confirm the compounds involved and to elucidate possible genotoxic effects of RAs.
- Klíčová slova
- algae, chemical analysis, complex mixtures, cyanobacteria, cyanotoxins, cytotoxicity, extracts, genotoxicity, microcystins, retinoic acids,
- MeSH
- alkaloidy izolace a purifikace toxicita MeSH
- buňky Hep G2 MeSH
- Chlorophyta metabolismus MeSH
- chromatografie kapalinová MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- kometový test MeSH
- lidé MeSH
- mikrocystiny izolace a purifikace toxicita MeSH
- mikrojaderné testy MeSH
- mikrojádra chromozomálně defektní chemicky indukované MeSH
- poškození DNA * MeSH
- sinice metabolismus MeSH
- tandemová hmotnostní spektrometrie MeSH
- toxiny kmene Cyanobacteria MeSH
- tretinoin izolace a purifikace toxicita MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alkaloidy MeSH
- cylindrospermopsin MeSH Prohlížeč
- microcystin MeSH Prohlížeč
- mikrocystiny MeSH
- toxiny kmene Cyanobacteria MeSH
- tretinoin MeSH
