Constitutive nuclear factor-kappa B mRNA, protein overexpression and enhanced DNA-binding activity in thymidylate synthase inhibitor-resistant tumour cells.
In this study, the gene copy number, mRNA and protein expression levels and nuclear DNA-binding activity of nuclear factor kappa B (NF-kappa B) were compared in a panel of five pairs of thymidylate synthase (TS) inhibitor-resistant and wild-type parent cancer cell lines. High constitutive NF-kappa B DNA-binding activity was detected in all chemoresistant cell lines. The upregulated NF-kappa B activity was composed of NF-kappa B subunits p50 and p65. Four out of five resistant cell lines constitutively overexpressed NF-kappa B p50 and p65 mRNA and protein. One resistant cell line with the highest NF-kappa B DNA-binding activity showed normal p50 and p65 protein expression. No NF-kappa B gene amplification was detected in resistant cell lines. Transient exposure of wild-type RKO(WT) and H630(WT) cells to 5-FU induced NF-kappa B DNA-binding activity but had no effect on NF-kappa B protein expression in these cells. Our results indicate that high constitutive NF-kappa B activity caused by its gene overexpression is an intrinsic character of TS inhibitor-resistant cells. NF-kappa B can antagonise anticancer drug-induced apoptosis. High NF-kappa B expression and nuclear activity in TS inhibitor-resistant cancer cells may play an important role in the chemoresistance.
Wang
W
W
Department of Medicine and Therapeutics, Institute of Medical Sciences, University of Aberdeen, Foresterhill, UK. w.wang@beatson.gla.ac.uk
Cassidy
J
J
D002455:Cell Division / Q000187:drug effects; D004247:DNA / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm; D024202:Electrophoretic Mobility Shift Assay; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005472:Fluorouracil / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit; D011485:Protein Binding / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors*; D051996:Transcription Factor RelA; D014407:Tumor Cells, Cultured
D004791:Enzyme Inhibitors; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D012333:RNA, Messenger; D051996:Transcription Factor RelA; D004247:DNA; D013940:Thymidylate Synthase; D005472:Fluorouracil