Nuclear factor-kappa B is upregulated in colorectal cancer

Surgery. 2001 Aug;130(2):363-9. doi: 10.1067/msy.2001.116672.

Abstract

Background: Chemoresistance may involve the anti-apoptotic transcriptional regulator, nuclear factor-kappa B (NF-kappa B). The purpose of this study was to determine whether chemotherapy induces NF-kappa B activation in a human colon cancer cell line (SW48) and whether NF-kappa B is constitutively activated in colorectal cancer.

Methods: SW48 cells were incubated with gemcitabine hydrochloride (Gemzar) in the presence and absence of the 26s proteasome inhibitor, MG132, and NF-kappa B binding (electrophoretic mobility shift assay), DNA synthesis (tritiated thymidine uptake), cell viability (3-[4,5-dimethylthiazol-2-yl]-diphenyl-tetrazolium bromide assay), and apoptosis (caspase-3 activity) were measured at 24 hours. NF-kappa B binding (electrophoretic mobility shift assay) was also assayed in 10 colorectal cancer tumors.

Results: SW48 cells demonstrated constitutive NF-kappa B binding that was enhanced by gemcitabine hydrochloride in a dose-dependent manner. MG132 inhibited NF-kappa B binding and enhanced gemcitabine hydrochloride's inhibition of DNA synthesis (gemcitabine hydrochloride = 73% +/- 1.4% vs gemcitabine hydrochloride + MG132 = 6% +/- 0.4%, P <.05), cell killing (gemcitabine hydrochloride = 87% +/- 2.0 vs gemcitabine hydrochloride + MG132 = 25% +/- 1.3%, P <.05), and caspase-3 activity (gemcitabine hydrochloride = 870 +/- 17.4 vs gemcitabine hydrochloride + MG132 = 1075 +/- 20.4, P <.05). NF-kappa B binding was increased in 8 of 10 colorectal cancer tumors compared with adjacent normal mucosa.

Conclusions: Gemcitabine hydrochloride enhances NF-kappa B binding in a colorectal cancer cell line, whereas inhibition of NF-kappa B enhances gemcitabine hydrochloride's antitumor activity. NF-kappa B is also activated in human colorectal cancer. NF-kappa B may identify chemoresistant tumors, whereas inhibition of NF-kappa B may be a novel, biologically based therapy. (Surgery 2001;130:363-9).

MeSH terms

  • Antimetabolites, Antineoplastic / toxicity
  • Caspase 3
  • Caspases / metabolism
  • Colon / metabolism
  • Colon / pathology
  • Colorectal Neoplasms / metabolism
  • Colorectal Neoplasms / pathology*
  • Cysteine Proteinase Inhibitors / pharmacology
  • DNA / biosynthesis
  • Deoxycytidine / analogs & derivatives
  • Deoxycytidine / toxicity
  • Humans
  • Intestinal Mucosa / metabolism
  • Intestinal Mucosa / pathology
  • Leupeptins / pharmacology
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / metabolism*
  • Protein Binding / drug effects
  • Protein Binding / physiology
  • Rectum / metabolism
  • Rectum / pathology
  • Tumor Cells, Cultured

Substances

  • Antimetabolites, Antineoplastic
  • Cysteine Proteinase Inhibitors
  • Leupeptins
  • NF-kappa B
  • Deoxycytidine
  • DNA
  • gemcitabine
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde