Role of Rel/NF‐κB transcription factors in apoptosis of human hepatocellular carcinoma cells

BACKGROUND Primary hepatocellular carcinoma (HCC) is one of the 10 most common human carcinomas in the world. The mechanism by which HCC cells resist apoptosis induced by various treatment modalities is poorly understood. METHODS The authors sought to determine whether Rel/NF‐κB transcription factors play a key role in controlling apoptosis in human HCCcells. We assessed constitutive and inducible activation of NF‐κB in hepatitis B virus (HBV)‐positive (Hep3B) and in hepatitis virus‐negative (Chang, HepG2) HCC cells, as well as the role of known inhibitors of NF‐κB activity. RESULTS The current study data demonstrate that 1) RelA/NF‐κB activity is activated constitutively in Hep3B cells, as determined by electrophoretic mobility shift assays; 2) RelA/NF‐κB reporter gene activity is inhibited specifically by dominant‐negative mutants of IκBα, IKK1, IKK2, MEKK1, and MEKK3 and it is activated by overexpression of wild‐type MEKK3, suggesting that upstream kinase cascades induce phosphorylation of IκBα and activate RelA/NF‐κB in Hep3B cells; 3) overexpression of the HBV x gene fails to activate NF‐κB in HepG2 and Chang cell lines; 4) The NF‐κB–inducible gene, bcl‐xl, is overexpressed in Hep3B cells and is inhibited by the proteosome inhibitor PS341, which prevents IκBα degradation and RelA/NF‐κB activation; and 5) inhibition of constitutive RelA/NF‐κB activity by PS341 sensitizes Hep3B cells to doxorubicin‐induced apoptosis. CONCLUSIONS These results are consistent with the role of RelA/NF‐κB activity in the regulation of apoptosis through activation of its downstream target genes and suggest that signaling pathways that control RelA/NF‐κB activity may be important targets for novel therapeutic approaches in the treatment of human HCC. Cancer 2002;95:1696–705. © 2002 American Cancer Society. DOI 10.1002/cncr.10829