Expression of Nuclear Factor-κB and Its Clinical Significance in Nonsmall-Cell Lung Cancer

doi:10.1016/j.athoracsur.2006.01.049

The Annals of Thoracic Surgery

Volume 82, Issue 1, July 2006, Pages 243-248

https://doi.org/10.1016/j.athoracsur.2006.01.049 Get rights and content

Background

It has been known that transcription factor nuclear factor (NF)-κB plays an important role in cell proliferation and oncogenesis. The aims of this study were to evaluate expression levels of NF-κB in nonsmall-cell lung cancer (NSCLC) and to elucidate its clinical significance and prognostic value for patients with NSCLC.

Methods

Using 45 tumor tissue specimens from 45 patients with NSCLC who underwent surgery, we investigated the expression of NF-κB using Western blotting analysis. Apoptotic rate of NSCLC cells with different expression of NF-κB was determined by TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end-labeling) assay. Paraffin-embedded tissue blocks from 71 consecutive patients with NSCLC were obtained for immunohistochemical staining.

Results

The expression level of NF-κB in poorly or moderately differentiated lung cancer cells was higher than that in well-differentiated ones (p = 0.001). The apoptotic rate was lower for lung cancer cells with higher NF-κB expression than for those with lower NF-κB expression (p = 0.0238). Furthermore, expression of NF-κB was correlated with caspase-3, cyclooxygenase-2, and p53 expression in lung cancer cells that were examined. Most NSCLC cells showed nuclear staining pattern and the nuclear positive rate was 67.6% (48 of 71 specimens). Immunohistochemical NF-κB expression in patients with NSCLC was an independent prognostic factor for overall survival.

Conclusions

The results of this study suggest that expression of NF-κB may correlate with lung cancer differentiation. Overexpression of NF-κB inhibits tumor cell apoptosis and indicates an unfavorable prognosis for overall survival in some patients with NSCLC.

Section snippets

Patients

Forty-five tumor tissue specimens were sampled from 45 patients with NSCLC who underwent surgery between October and December 2004. All patients were required to sign a written informed consent. Patients who received induction therapy were excluded from the study. Samples were histologically confirmed to have been obtained from tumor masses. A portion of each sample was frozen immediately after surgical resection and stored at –80°C until use. The clinicopathologic features of the specimens

Correlation With Clinical Characteristics

The Western blotting products from NF-κB protein were semiquantified using β-actin as an intrinsic control. The density of each band was separately converted to a numeric value by Bandleader software. The ratio of NF-κB to β-actin in each case was calculated. The average NF-κB/β-actin value was 0.6047 ± 0.3572 (Fig 1).

There was no significant correlation between NF-κB and patient age, sex, tumor staging, and pathologic type. Expression level of NF-κB was higher in poorly and moderately

Comment

The transcription factor NF-κB has been implicated in the control of cell proliferation and oncogenesis [6]. Nuclear factor-κB transcription factors bind to DNA as heterodimers or homodimers that are composed of five possible subunits in mouse and human (RelA/p65, c-Rel, RelB, p50, and p52). These proteins mediate DNA binding, dimerization, and interactions with inhibitory factors known as inhibitor κB (I-κB) proteins [7]. Degradation of the I-κB proteins results in the liberation of NF-κB,

References (19)

C.F. Mountain et al.
Regional lymph node classification for lung cancer staging
Chest
(1997)
R.Z. Orlowski et al.
NF-kappaB as a therapeutic target in cancer
Trends Mol Med
(2002)
C.E. Denlinger et al.
Modulation of antiapoptotic cell signaling pathways in non-small cell lung cancerthe role of NF-kappaB
Semin Thorac Cardiovasc Surg
(2004)
K.H. Kang et al.
Caspase-3-mediated cleavage of the NF-kappa B subunit p65 at the NH2 terminus potentiates naphthoquinone analog-induced apoptosis
J Biol Chem
(2001)
H. Wu et al.
NF-kappa B activation of p53A potential mechanism for suppressing cell growth in response to stress
J Biol Chem
(1994)
A.B. Sandler et al.
COX-2 inhibition and lung cancer
Semin Oncol
(2004)
J.M. Yoon et al.
Adenovirus-uteroglobin suppresses COX-2 expression via inhibition of NF-kappaB activity in lung cancer cells
Lung Cancer
(2005)
O.P. Veiby et al.
Chemoresistanceimpact of nuclear factor (NF)-kappaB inhibition by small interfering RNA
Clin Cancer Res
(2004)
W.D. Travis et al.
Histological typing of lung and pleural tumorsInternational histological classification of tumors
(1999)

There are more references available in the full text version of this article.

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Original articleGeneral thoracicExpression of Nuclear Factor-κB and Its Clinical Significance in Nonsmall-Cell Lung Cancer

Background

Methods

Results

Conclusions

Section snippets

Patients

Correlation With Clinical Characteristics

Comment

Chest

Trends Mol Med

Semin Thorac Cardiovasc Surg

J Biol Chem

J Biol Chem

Semin Oncol

Lung Cancer

Chemoresistanceimpact of nuclear factor (NF)-kappaB inhibition by small interfering RNA

Clin Cancer Res

Histological typing of lung and pleural tumorsInternational histological classification of tumors

Original article
General thoracic
Expression of Nuclear Factor-κB and Its Clinical Significance in Nonsmall-Cell Lung Cancer