Antioxidant enzyme activities and expression in different metastatic B16 melanoma cell subsets.

<p>(A) and (C) Enzyme activities were measured in metastatic cell subsets isolated from growing liver or lung foci 7 days after inoculation and cultured for 48 h. The enzyme activities before inoculation in B16-F10 cells cultured for 48 h were: SOD1, 1.51±0.33 units/10⁶cells; SOD2, 0.14±0.05 units/10⁶cells; CAT, 4.22±1.05 milliunits/10⁶cells; GPX, 7.51±1.63 milliunits/10⁶cells; GR, 6.58±2.04 milliunits/10⁶cells; and NOX, 183±42 RLU/10⁶cells (n = 6 in all cases). iB16 cells were transfected <i>in vitro</i> with anti-Nrf2-siRNA as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096466#pone-0096466-t001" target="_blank">Table 1</a>. RLU, relative light units. Data are mean values ± S.D. (n = 5–6 in all cases). *p<0.05, **p<0.01 <i>versus</i> iB16 controls. Enzyme activities measured in iB16 cells transfected with Nrf2 sense or scrambled oligonucleotides were not significantly different from control values (not shown). (B) and (D)Melanoma cells isolated from liver or lung metastatic foci 7 days after inoculation were cultured for 48 h. Results obtained in iB16 cells transfected with lentiviral vector not harboring any gene (negative control) were not different from control values (not shown).Data from quantitative RT-PCR are expressed as mean fold change ± S.D. (n = 6 in all cases). *p<0.05, **p<0.01 <i>versus</i> iB16 controls. Enzyme expression measured in iB16 cells transfected with Nrf2 sense or scrambled oligonucleotides was not significantly different from control values (not shown).