Effects of CD30 activation on NF-κB activity.
(A) NF-κB activity determined by the gel-shift assays. Karpas 299 cell line was incubated with HeFi-1 or isotype-specific control IgG for 1 hour. NF-κB activity was determined by mobility shift observed in the gel electrophoresis of nuclear extracts. The left lanes in individual figures are loaded with radioactive labelled oligoprobes representing NF-κB binding sites and nuclear extracts (H). The right lanes are loaded with radioactive probes plus excess amounts of (× 200) unlabeled probes. These lanes represent the negative controls for the assays and characterize which bands are specific for NF-κB binding activity (C). In the Karpas 299 cell line, there was no NF-κB activity after control IgG incubation; CD30 activation caused activation of NF-κB. Jurkat cell line nuclear extracts were used as controls. N is the negative control, and P is the positive control. (B) Effects of the NF-κB inhibitor SN50 on systemic ALCL and Hodgkin lymphoma cell lines. Proliferation assays were performed in the presence of HeFi-1 and the NF-κB inhibitor SN50 on the Karpas 299 and KMH2 cell lines. SN50 alone caused inhibition of the KMH2 cell line, whereas it had no effect on the Karpas 299 cell line. Combined treatment with HeFi-1 and SN50 enhanced the inhibitory effects of HeFi-1 on the Karpas 299 cell line but had no significant additional effect on the KMH2 cell line. The results are presented as percentages of 3H-thymidine incorporation values compared to controls (IgG incubated). Each of 3 experiments was done in triplicate. A representative experiment is shown.
