NF-kappaB and apoptosis in colorectal tumourigenesis.
BACKGROUND
Nuclear factor-kappaB (NF-kappaB) may play an important role in colorectal tumourigenesis, controlling cell cycle and apoptosis gene expression. In addition, imbalances between cell proliferation and cell death are thought to underlie neoplastic development. The aims of this study were to investigate apoptosis and expression of several apoptosis-related proteins, and to determine correlations with colorectal tumour progression.


METHODS
Apoptosis was evaluated by the TUNEL assay in 48 patient samples, including adenomas, adenocarcinomas and adjacent normal mucosas. Immunohistochemistry was performed for Bcl-2 and NF-kappaB. Expression levels of p53, Bax and IkappaB proteins were determined by immunoblotting. Cultured human colon cancer cells were used to evaluate NF-kappaB expression and nuclear translocation by immunocytochemistry and immunoblotting.


RESULTS
Apoptosis and NF-kappaB immunoreactivity were significantly higher in tumour tissue compared with normal mucosa (P < 0.01), increasing in association with histological tumour progression (P < 0.01). Bcl-2 was consistently higher in normal mucosa (P < 0.01) and inversely correlated with the percentage of apoptosis (P < 0.01). Phosphorylated p53 and Bax levels were similar in tumour tissue and normal mucosa; however, the NF-kappaB inhibitor, IkappaB, tended to decrease in tumours. In vitro, nuclear translocation of NF-kappaB was greater in proliferative than in resting phases of colon cancer cells.


CONCLUSIONS
NF-kappaB expression and apoptosis are increased from adenoma to poorly differentiated adenocarcinoma tissues. Apoptosis is correlated with suppression of Bcl-2 expression, but appears to proceed through a p53- and Bax-independent pathway. Activation of NF-kappaB may play an important role in colorectal tumour progression.
Aranha
M M
MM
Centro de Patogénese Molecular, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal.
Borralho
P M
PM
Ravasco
P
P
Moreira da Silva
I B
IB
Correia
L
L
Fernandes
A
A
Camilo
M E
ME
Rodrigues
C M P
CM
D000236:Adenoma / Q000503:physiopathology*; D000368:Aged; D000369:Aged, 80 and over; D017209:Apoptosis; D014408:Biomarkers, Tumor / Q000378:metabolism*; D015179:Colorectal Neoplasms / Q000503:physiopathology*; D006801:Humans; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000378:metabolism*; D051020:bcl-X Protein / Q000378:metabolism*
D014408:Biomarkers, Tumor; D016328:NF-kappa B; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein