OBJECTIVES: Chemotherapeutic drugs induce senescence in cancer cells but, unlike replicative senescence or oncogene-induced senescence, do so rather inefficiently and depending on DNA damage. A thorough understanding of the biology of chemotherapy-induced senescent cells requires their isolation from a mixed population of adjacent senescent and non-senescent cancer cells. MATERIALS AND METHODS: We have developed and optimized a rapid iodixanol (OptiPrep)-based gradient centrifugation system to identify, isolate and characterize doxorubicin (DXR)-induced senescent hepatocellular carcinoma (HCC) cells (HepG2 and Huh-7) in vitro. RESULTS: After cellular exposure to DXR, we used iodixanol gradient-based centrifugation to isolate and re-plate cells on collagen-coated flasks, despite their low or null proliferative capacity. The isolated cell populations were enriched for DXR-induced senescent HCC cells, as confirmed by proliferation arrest assay, and β-galactosidase and DNA damage-dependent γH2A.X staining. CONCLUSIONS: Analysing pure cultures of chemotherapy-induced senescent versus non-responsive cancer cells will increase our knowledge on chemotherapeutic mechanisms of action, and help refine current therapeutic strategies.

• MeSH
• doxorubicin farmakologie   MeSH
• hepatocelulární karcinom patologie   MeSH
• kyseliny trijodbenzoové farmakologie   MeSH
• lidé MeSH
• nádory jater patologie   MeSH
• poškození DNA účinky léků   MeSH
• separace buněk * metody   MeSH
• stárnutí buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH