Background. Extracts from plant and/or animal tissues are frequently used in alternative medicine as drugs or food supplements. Such extracts may contain a complex of pharmacologically or physiologically active factors but frequently there exists no experimental confirmation as to precise mechanisms of action. This work aimed to verify if a long used bovine tissue extract Imuregen registered as a food supplement has desirable effect on tumor cells. Methods. Two independent methodological approaches were used. Viability of cell cultures was evaluated using WST-1-based cell cytotoxicity assay. Cell growth was monitored in real time using xCELLigence cell analysis. Normal human adherent lung fibroblasts (NHLF) were used to represent non - tumor lung cells. A human non-small cell lung carcinoma cell line H1299 was used as a model of tumor cells. Results. Our study demonstrated a direct influence on viability of the H1299 tumor cell line (p < 0.005) and a cytostatic/cytotoxic effect of the bovine tissue extract after 72h. of cultivation while leaving non-tumor NHLF cell line unaffected. The extract (0.1 μg/ml and 1 μg/ml, resp.) also significantly affected the viability of irradiated H1299 tumor cell line (p < 0.005, Co, 4Gy) compared to non-tumor irradiated counterparts. In addition to the cytotoxic effect, the extract slightly modified the generation time of the cells and substantial differences between the effects on tumor and non-tumor cell lines were observed. Conclusion. The data presented here might suggest the extract intervenes into the proliferative cell cycle and subsequently influences the generation time of cells. Further analyses should be oriented toward the effects of animal tissue extracts on cellular systems defending against tumors and/or infections and intercellular communications that lead to influencing the fate of individual cell types.

• Klíčová slova
• Imuregen,
• MeSH
• nádorové buněčné linie imunologie   účinky léků   MeSH
• nemalobuněčný karcinom plic genetika   imunologie   MeSH
• oncogene addiction MeSH
• proliferace buněk genetika   účinky léků   účinky záření   MeSH
• protinádorové látky imunologicky aktivní * MeSH
• techniky in vitro MeSH
• tkáňové extrakty imunologie   terapeutické užití   MeSH
• Publikační typ
• práce podpořená grantem MeSH

The aim of this study was to compare the effects of DNA repair inhibitors in the context of radio-sensitization of human lung cells. The radio-sensitizing effects of NU7441 (1 mM), an inhibitor of DNA-dependent protein kinase (DNA-PK); KU55933 (10 μM), an inhibitor of ataxia-telangiectasia mutated kinase (ATM); and VE-821 (10 μM), an inhibitor of ATM-related kinase (ATR) were tested by the xCELLigence system for monitoring proliferation, fluorescence microscopy for DNA damage detection, flow-cytometry for cell cycle and apoptosis analysis and western blotting and ELISA for determination of DNA repair proteins. We employed normal human lung fibroblasts (NHLF, p53-wild-type) and non-small cell lung cancer cells (H1299, p53-negative). DNA-PK inhibition (by NU7441) in combination with ionizing radiation (IR) increased the number of double strand breaks (DSB), which persisted 72 h after irradiation in both cell lines. Additionally, NU7441 and KU55933 in combination with IR caused G2-arrest. ATR inhibitor (VE-821) together with IR markedly inhibited proliferation and induced G2/M arrest accompanied by apoptosis in H1299, but not in NHLF cells, and thus diminished DNA-repair of tumour cells but not normal lung fibroblasts. Our findings indicate that ATR inhibition could be a promising therapeutic strategy in p53-deficient lung tumours.

• MeSH
• enzymy opravy DNA antagonisté a inhibitory   genetika   MeSH
• fibroblasty účinky záření   MeSH
• kultivované buňky účinky záření   MeSH
• nádorové buňky kultivované účinky léků   účinky záření   MeSH
• nemalobuněčný karcinom plic genetika   radioterapie   MeSH
• oprava DNA účinky léků   MeSH
• proliferace buněk účinky záření   MeSH
• radiosenzibilizující látky farmakologie   izolace a purifikace   terapeutické užití   MeSH
• techniky in vitro MeSH
• Publikační typ
• práce podpořená grantem MeSH