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Comprehensive Evaluation of Nuclear Factor-κΒ Expression Patterns in Non-Small Cell Lung Cancer.

Giopanou I, Lilis I, Papaleonidopoulos V, Marazioti A, Spella M, Vreka M, Papadaki H, Stathopoulos GT - PLoS ONE (2015)

Bottom Line: We found that the expression of the different NF-κB subunits was not concordant, warranting our integral approach.Overall, RelA, RelB, and P50 were expressed at higher levels compared with P52/P100.We conclude that pathologic studies of NF-κB expression in cancer should include multiple pathway components.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Molecular Respiratory Carcinogenesis, Department of Physiology, Faculty of Medicine, University of Patras, Rio, Achaia, Greece.

ABSTRACT
Nuclear factor (NF)-κB signalling is required for lung adenocarcinoma development in mice, and both of its subunits RelA and RelB were independently reported to be highly expressed in human non-small cell lung cancer (NSCLC). To comprehensively examine NF-κB expression in NSCLC, we analyzed serial sections of primary tumor samples from 77 well-documented patients (36 adenocarcinomas, 40 squamous cell carcinomas and 3 large cell carcinomas) for immunoreactivity of RelA, RelB, P50, and P52/P100. Tumor and intratumoral stroma areas were discriminated based on proliferating cell nuclear antigen immunoreactivity and inflammatory infiltration was assessed in intratumoral stroma areas. NF-κB immunoreactivity was quantified by intensity, extent, and nuclear localization and was cross-examined with tumor cell proliferation, inflammatory infiltration, and clinical-pathologic data. We found that the expression of the different NF-κB subunits was not concordant, warranting our integral approach. Overall, RelA, RelB, and P50 were expressed at higher levels compared with P52/P100. However, RelA and P50 were predominantly expressed in intratumoral stroma, but RelB in tumor cells. Importantly, tumor area RelA expression was correlated with the intensity of inflammatory infiltration, whereas RelB expression was identified in proliferating tumor cells. Using multiple logistic regression, we identified that tumor RelB expression was an independent predictor of lymph node metastasis, and tumor P50 was an independent predictor of TNM6 stage IIB or higher, whereas tumor RelA was an independent predictor of inflammatory infiltration. We conclude that pathologic studies of NF-κB expression in cancer should include multiple pathway components. Utilizing such an approach, we identified intriguing associations between distinct NF-κB subunits and clinical and pathologic features of NSCLC.

No MeSH data available.


Related in: MedlinePlus

Association of NF-κB expression with clinical and pathologic parameters in 77 patients with NSCLC.(A) NF-κB expression levels subdivided by clinical and pathological parameters. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, *, and **: P > 0.05, P < 0.05, and P < 0.0501 for indicated comparisons by Wilcoxon signed rank tests or Kruskal-Wallis tests followed by Dunn’s post-tests, for two or multiple comparison groups, respectively. (B) Results of binary logistic regression analyses using NF-κB subunit expression scores as the input (independent variables) and dichotomized clinical and pathologic parameters as the output (dependent variables). RR, risk ratios; CI, confidence intervals; P, probability values.
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pone.0132527.g004: Association of NF-κB expression with clinical and pathologic parameters in 77 patients with NSCLC.(A) NF-κB expression levels subdivided by clinical and pathological parameters. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, *, and **: P > 0.05, P < 0.05, and P < 0.0501 for indicated comparisons by Wilcoxon signed rank tests or Kruskal-Wallis tests followed by Dunn’s post-tests, for two or multiple comparison groups, respectively. (B) Results of binary logistic regression analyses using NF-κB subunit expression scores as the input (independent variables) and dichotomized clinical and pathologic parameters as the output (dependent variables). RR, risk ratios; CI, confidence intervals; P, probability values.

Mentions: We next examined whether NF-κB subunit expression in tumor and stroma compartments is correlated with clinical and pathologic features of our patients with NSCLC. For this, correlation analyses were done, which revealed no significant associations (data not shown). NF-κB scores were further subdivided according to clinical and pathologic features and subgroups were compared (Fig 4A). The stroma score of P100/P52 was significantly lower in well-differentiated tumors, as compared with low or moderately differentiated ones. In addition, stromal P100/P52 expression was decreased in patients with N2 disease, as compared with patients without lymphatic involvement. No other significant differences were identified regarding the intratumoral stromal expression of any other subunit. When tumor area NF-κB scores were examined this way, RelA was found to be expressed at higher levels in men compared with women, in patients aged 65 and over compared with younger ones, and in patients with squamous cell carcinoma compared with adenocarcinoma. Importantly, tumor P50 scores were elevated in patients with N2 and pTNM6 stage III disease compared with patients without lymphatic involvement and stage II disease, respectively. Finally, tumor P100/P52 expression was statistically significantly increased in patients with pTNM6 stage II disease compared with patients with stage I disease. In a third level of investigations, clinical and pathologic NSCLC features were dichotomized by their median and were entered as dependents into binary logistic regression analyses, using tumor and stromal NF-κB scores as the input (independent) variables (Fig 4B). These analyses revealed that tumor RelA score was an independent predictor of age 65 or older, of squamous histology, and of inflammatory infiltration. Interestingly, tumor and stroma RelB expression were identified as positive and negative predictors of nodal involvement, respectively. Finally, tumor P50 score was found to be a independent predictor of poor differentiation and pTNM6 stage IIB or higher. Hence distinct NF-κB subunit (RelB and P50, but not RelA) expression was associated with advanced disease, and distinct NF-κB subunits were linked with defined features of NSCLC, validating our integrated approach.


Comprehensive Evaluation of Nuclear Factor-κΒ Expression Patterns in Non-Small Cell Lung Cancer.

Giopanou I, Lilis I, Papaleonidopoulos V, Marazioti A, Spella M, Vreka M, Papadaki H, Stathopoulos GT - PLoS ONE (2015)

Association of NF-κB expression with clinical and pathologic parameters in 77 patients with NSCLC.(A) NF-κB expression levels subdivided by clinical and pathological parameters. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, *, and **: P > 0.05, P < 0.05, and P < 0.0501 for indicated comparisons by Wilcoxon signed rank tests or Kruskal-Wallis tests followed by Dunn’s post-tests, for two or multiple comparison groups, respectively. (B) Results of binary logistic regression analyses using NF-κB subunit expression scores as the input (independent variables) and dichotomized clinical and pathologic parameters as the output (dependent variables). RR, risk ratios; CI, confidence intervals; P, probability values.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4493092&req=5

pone.0132527.g004: Association of NF-κB expression with clinical and pathologic parameters in 77 patients with NSCLC.(A) NF-κB expression levels subdivided by clinical and pathological parameters. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, *, and **: P > 0.05, P < 0.05, and P < 0.0501 for indicated comparisons by Wilcoxon signed rank tests or Kruskal-Wallis tests followed by Dunn’s post-tests, for two or multiple comparison groups, respectively. (B) Results of binary logistic regression analyses using NF-κB subunit expression scores as the input (independent variables) and dichotomized clinical and pathologic parameters as the output (dependent variables). RR, risk ratios; CI, confidence intervals; P, probability values.
Mentions: We next examined whether NF-κB subunit expression in tumor and stroma compartments is correlated with clinical and pathologic features of our patients with NSCLC. For this, correlation analyses were done, which revealed no significant associations (data not shown). NF-κB scores were further subdivided according to clinical and pathologic features and subgroups were compared (Fig 4A). The stroma score of P100/P52 was significantly lower in well-differentiated tumors, as compared with low or moderately differentiated ones. In addition, stromal P100/P52 expression was decreased in patients with N2 disease, as compared with patients without lymphatic involvement. No other significant differences were identified regarding the intratumoral stromal expression of any other subunit. When tumor area NF-κB scores were examined this way, RelA was found to be expressed at higher levels in men compared with women, in patients aged 65 and over compared with younger ones, and in patients with squamous cell carcinoma compared with adenocarcinoma. Importantly, tumor P50 scores were elevated in patients with N2 and pTNM6 stage III disease compared with patients without lymphatic involvement and stage II disease, respectively. Finally, tumor P100/P52 expression was statistically significantly increased in patients with pTNM6 stage II disease compared with patients with stage I disease. In a third level of investigations, clinical and pathologic NSCLC features were dichotomized by their median and were entered as dependents into binary logistic regression analyses, using tumor and stromal NF-κB scores as the input (independent) variables (Fig 4B). These analyses revealed that tumor RelA score was an independent predictor of age 65 or older, of squamous histology, and of inflammatory infiltration. Interestingly, tumor and stroma RelB expression were identified as positive and negative predictors of nodal involvement, respectively. Finally, tumor P50 score was found to be a independent predictor of poor differentiation and pTNM6 stage IIB or higher. Hence distinct NF-κB subunit (RelB and P50, but not RelA) expression was associated with advanced disease, and distinct NF-κB subunits were linked with defined features of NSCLC, validating our integrated approach.

Bottom Line: We found that the expression of the different NF-κB subunits was not concordant, warranting our integral approach.Overall, RelA, RelB, and P50 were expressed at higher levels compared with P52/P100.We conclude that pathologic studies of NF-κB expression in cancer should include multiple pathway components.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Molecular Respiratory Carcinogenesis, Department of Physiology, Faculty of Medicine, University of Patras, Rio, Achaia, Greece.

ABSTRACT
Nuclear factor (NF)-κB signalling is required for lung adenocarcinoma development in mice, and both of its subunits RelA and RelB were independently reported to be highly expressed in human non-small cell lung cancer (NSCLC). To comprehensively examine NF-κB expression in NSCLC, we analyzed serial sections of primary tumor samples from 77 well-documented patients (36 adenocarcinomas, 40 squamous cell carcinomas and 3 large cell carcinomas) for immunoreactivity of RelA, RelB, P50, and P52/P100. Tumor and intratumoral stroma areas were discriminated based on proliferating cell nuclear antigen immunoreactivity and inflammatory infiltration was assessed in intratumoral stroma areas. NF-κB immunoreactivity was quantified by intensity, extent, and nuclear localization and was cross-examined with tumor cell proliferation, inflammatory infiltration, and clinical-pathologic data. We found that the expression of the different NF-κB subunits was not concordant, warranting our integral approach. Overall, RelA, RelB, and P50 were expressed at higher levels compared with P52/P100. However, RelA and P50 were predominantly expressed in intratumoral stroma, but RelB in tumor cells. Importantly, tumor area RelA expression was correlated with the intensity of inflammatory infiltration, whereas RelB expression was identified in proliferating tumor cells. Using multiple logistic regression, we identified that tumor RelB expression was an independent predictor of lymph node metastasis, and tumor P50 was an independent predictor of TNM6 stage IIB or higher, whereas tumor RelA was an independent predictor of inflammatory infiltration. We conclude that pathologic studies of NF-κB expression in cancer should include multiple pathway components. Utilizing such an approach, we identified intriguing associations between distinct NF-κB subunits and clinical and pathologic features of NSCLC.

No MeSH data available.


Related in: MedlinePlus