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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

Immunohistochemical detection of NF-κB subunits in NSCLC, juxta-tumoral normal lung structures and preneoplastic lesions.(A) Representative images. Images in red frames representatively display differential NF-κB subunit expression in tumor and intratumoral stroma areas. (B) Overall scoring of NF-κB subunit expression levels. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, * and ***: P > 0.05, P < 0.05, and P < 0.001 for indicated comparisons by Friedman’s test followed by Dunn’s post-tests. (C) Co-expression matrixes of categorical NF-κB subunit expression levels. For this, NF-κB scores from (B) were categorized into low (0–4), intermediate (5–6), and high (7–18). ns: P > 0.05 by χ2 tests followed by Fisher’s exact tests.
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pone.0132527.g001: Immunohistochemical detection of NF-κB subunits in NSCLC, juxta-tumoral normal lung structures and preneoplastic lesions.(A) Representative images. Images in red frames representatively display differential NF-κB subunit expression in tumor and intratumoral stroma areas. (B) Overall scoring of NF-κB subunit expression levels. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, * and ***: P > 0.05, P < 0.05, and P < 0.001 for indicated comparisons by Friedman’s test followed by Dunn’s post-tests. (C) Co-expression matrixes of categorical NF-κB subunit expression levels. For this, NF-κB scores from (B) were categorized into low (0–4), intermediate (5–6), and high (7–18). ns: P > 0.05 by χ2 tests followed by Fisher’s exact tests.

Mentions: The clinical and pathologic features of the study patients are summarized in Table 1. The raw study results are appended to this article as S1 Table. We first examined NF-κΒ subunit expression in normal lung areas adjacent to our surgically resected tumor samples. In bronchial and alveolar epithelium, RelA, P50 and P100/P52 exhibited low or moderate cytoplasmic immunoreactivity, while RelB displayed higher expression levels. In juxta-tumoral bronchial and alveolar hyperplasias, immunoreactivity for all NF-κB subunits showed a stronger expression pattern, with RelB showing both cytoplasmic and nuclear localization. In tumors, all NF-κB subunits were highly expressed relative to normal and hyperplastic areas, with RelB scoring highest and showing the highest degree of nuclear localization. However, RelA, RelB, and P50 immunoreactivity was stronger compared with P100/P52 (Fig 1A and 1B). When NF-κB subunit scores were subdivided into low (0–4), intermediate (5–6), or high (7–18), no relationship was evident between the different subunits (Fig 1C). No statistically significant differences were noted between the different NSCLC histologic subtypes. However, statistically significant differences were observed in NF-κB subunit expression levels within each tumor: tumor areas displayed different expression patterns compared with intratumoral stroma areas (Fig 1A; representative panels in red frame). These results suggested that multiple NF-κΒ subunits are progressively overexpressed in NSCLC, showed that not all NF-κΒ subunits are expressed equally, and identified a discordance in the expression of multiple NF-κΒ subunits between tumor and intratumoral stromal areas, warranting further investigation.


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)

Immunohistochemical detection of NF-κB subunits in NSCLC, juxta-tumoral normal lung structures and preneoplastic lesions.(A) Representative images. Images in red frames representatively display differential NF-κB subunit expression in tumor and intratumoral stroma areas. (B) Overall scoring of NF-κB subunit expression levels. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, * and ***: P > 0.05, P < 0.05, and P < 0.001 for indicated comparisons by Friedman’s test followed by Dunn’s post-tests. (C) Co-expression matrixes of categorical NF-κB subunit expression levels. For this, NF-κB scores from (B) were categorized into low (0–4), intermediate (5–6), and high (7–18). ns: P > 0.05 by χ2 tests followed by Fisher’s exact tests.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0132527.g001: Immunohistochemical detection of NF-κB subunits in NSCLC, juxta-tumoral normal lung structures and preneoplastic lesions.(A) Representative images. Images in red frames representatively display differential NF-κB subunit expression in tumor and intratumoral stroma areas. (B) Overall scoring of NF-κB subunit expression levels. Data presented as median with boxes indicating interquartile range and whiskers indicating 95% percentiles. ns, * and ***: P > 0.05, P < 0.05, and P < 0.001 for indicated comparisons by Friedman’s test followed by Dunn’s post-tests. (C) Co-expression matrixes of categorical NF-κB subunit expression levels. For this, NF-κB scores from (B) were categorized into low (0–4), intermediate (5–6), and high (7–18). ns: P > 0.05 by χ2 tests followed by Fisher’s exact tests.
Mentions: The clinical and pathologic features of the study patients are summarized in Table 1. The raw study results are appended to this article as S1 Table. We first examined NF-κΒ subunit expression in normal lung areas adjacent to our surgically resected tumor samples. In bronchial and alveolar epithelium, RelA, P50 and P100/P52 exhibited low or moderate cytoplasmic immunoreactivity, while RelB displayed higher expression levels. In juxta-tumoral bronchial and alveolar hyperplasias, immunoreactivity for all NF-κB subunits showed a stronger expression pattern, with RelB showing both cytoplasmic and nuclear localization. In tumors, all NF-κB subunits were highly expressed relative to normal and hyperplastic areas, with RelB scoring highest and showing the highest degree of nuclear localization. However, RelA, RelB, and P50 immunoreactivity was stronger compared with P100/P52 (Fig 1A and 1B). When NF-κB subunit scores were subdivided into low (0–4), intermediate (5–6), or high (7–18), no relationship was evident between the different subunits (Fig 1C). No statistically significant differences were noted between the different NSCLC histologic subtypes. However, statistically significant differences were observed in NF-κB subunit expression levels within each tumor: tumor areas displayed different expression patterns compared with intratumoral stroma areas (Fig 1A; representative panels in red frame). These results suggested that multiple NF-κΒ subunits are progressively overexpressed in NSCLC, showed that not all NF-κΒ subunits are expressed equally, and identified a discordance in the expression of multiple NF-κΒ subunits between tumor and intratumoral stromal areas, warranting further investigation.

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