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. 2009 Sep 15;101(8):1290–1297. doi: 10.1038/sj.bjc.6605311

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Copyright © 2009 Cancer Research UK

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Hypoxia-inducible factor (HIF) activity correlated with oxaliplatin resistance in spheroid cultures. A HCT116 reporter cell line stably expressing a constitutive renilla luciferase and hypoxia-induced (LDH-HRE) firefly luciferase was used to validate the fractionation methodology. Hypoxia-inducible factor-1 activity was measured in sorted spheroid sub-populations by determining the relative expression of firefly to renilla luciferase (A). HCT116 EV (B) and DN (C) spheroids were fractionated on the basis of Hoechst 33342 fluorescence and the resulting sub-populations were exposed to various concentrations of oxaliplatin. Cell survival was assessed by clonogenic assay. Data are the mean of three experiments ±s.e.