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Modifications of the anti‐oxidant metabolism during proliferation and differentiation of colon tumor cell lines

The anti‐oxidant metabolism was studied at different times after sub‐culture in 2 colon cell lines previously characterized for their growth and differentiation properties. The HT29 cell line is mainly composed of proliferative and undifferentiative cells, while the derived 5‐fluorouracil (FUra)‐adapted cells undergo growth‐dependent differentiation, which is complete at post‐confluence. In the 2 cell lines, all the anti‐oxidant parameters studied appeared to be related to proliferation, with increased activity of superoxide dismutase (SOD) 1 and 2, catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GSR), and glutathione transferase (GST), and decreased glucose‐6‐phosphate dehydrogenase (G6PD) activity and glutathione content, in parallel with slowing down of proliferation. At post‐confluence, these metabolic parameters remained stable, except for GPX activity, which continued to increase, and CAT activity, which decreased. The amounts of SOD1, SOD2 and CAT immunoreactive proteins, estimated by Western blotting, appeared to be correlated to their respective enzymatic activities, SOD1, CAT and GST activity and glutathione content, which remained at similar levels in the 2 cell lines for all times studied, appeared unrelated to the differentiation process. GSR and GPX activity, which was lower in FUraadapted than in parental cells only at post‐confluence, could be considered as markers of differentiated cells. The higher SOD2 and lower G6PD activity observed in FUra‐resistant cell in comparison with parental cells at all times after sub‐culture could be characteristic both of differentiative and of differentiated cells. Interestingly, cytogenetics have previously indicated that deletions of the long arm of chromosome 6, which carry the gene for SOD2, were frequently observed in parental but not in FUra‐adapted cells. These results demonstrate that modifications of the anti‐oxidant metabolism occur in relation with proliferation and differentiation, and suggest a particular role for SOD2 in these cellular processes.