EGFR nuclear import in gallbladder carcinoma

Nuclear phosphorylated EGFR upregulates iNOS expression and confers independent prognostic impact

Chien Feng Li, Fu Ming Fang, Ju-Ming Wang, Ching Cherng Tzeng, Hui Chun Tai, Yu Ching Wei, Shau Hsuan Li, Yuan Ting Lee, Yu Hui Wang, Shih Chen Yu, Yow Ling Shiue, Patrick Yu Wei Chu, Wen Ling Wang, Li Tzong Chen, Hsuan Ying Huang

研究成果: Article

16 引文 (Scopus)

摘要

Background: The understanding of epidermal growth factor receptor (EGFR) deregulation in carcinogenesis remains incomplete. We investigated the implications of EGFR gene status and EGFR nuclear translocation in gallbladder carcinoma (GBCA). Methods: Subcellular localization of EGFR and phosphorylated EGFR (pEGFR) was analyzed by fractional immunoblotting and confocal immunofluorescence in GBCA cell lines. pEGFR binding to iNOS promoter was assessed by chromatin immunoprecipitation with iNOS promoter activity evaluated by luciferase assay. EGFR, pEGFR, and iNOS were immunohistochemically assessable for localization and level in the training set of 104 GBCAs on tissue microarrays, with 76 cases analyzed for EGFR gene by chromogenic in situ hybridization (CISH) and mutant-enriched PCR targeting exons 19 and 21. The prognostic impact of nuclear pEGFR (N-pEGFR) immunoexpression was reaffirmed on whole sections of 58 GBCAs in the test set. Results: Nuclear expression of EGFR and pEGFR was substantiated in vitro with augmented activity of iNOS promoter elicited by pEGFR binding upon EGF treatment. Despite no mutation, EGFR amplification, identified in 11 cases (15%) by CISH, strongly correlated with cytoplasmic EGFR expression (P < 0.001) but not with disease-specific survival (DSS). Immunoexpression of nuclear EGFR (N-EGFR), cytoplasmic pEGFR, and N-pEGFR was strongly related to that of iNOS (all ≤0.005). N-pEGFR independently predicted worse DSS in both training (P = 0.0468, HR = 2.024) and test sets (P = 0.0223, HR = 5.573). Conclusions: N-EGFR and N-pEGFR express in GBCA, conferring clinical aggressiveness partly through iNOS transactivation. Lacking response-predicting mutation, EGFR gene status, albeit amplified in 15% of GBCA, is neither related to nuclear EGFR translocation nor prognostically useful.

原文English
頁(從 - 到)443-454
頁數12
期刊Annals of Surgical Oncology
19
發行號2
DOIs
出版狀態Published - 2012 二月 1

指紋

Cell Nucleus Active Transport
Gallbladder
Epidermal Growth Factor Receptor
Up-Regulation
Carcinoma
erbB-1 Genes
In Situ Hybridization
Mutation
Chromatin Immunoprecipitation
Luciferases
Immunoblotting
Epidermal Growth Factor
Transcriptional Activation
Fluorescent Antibody Technique
Exons
Carcinogenesis
Cell Line
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Surgery
  • Oncology

引用此文

Li, Chien Feng ; Fang, Fu Ming ; Wang, Ju-Ming ; Tzeng, Ching Cherng ; Tai, Hui Chun ; Wei, Yu Ching ; Li, Shau Hsuan ; Lee, Yuan Ting ; Wang, Yu Hui ; Yu, Shih Chen ; Shiue, Yow Ling ; Chu, Patrick Yu Wei ; Wang, Wen Ling ; Chen, Li Tzong ; Huang, Hsuan Ying. / EGFR nuclear import in gallbladder carcinoma : Nuclear phosphorylated EGFR upregulates iNOS expression and confers independent prognostic impact. 於: Annals of Surgical Oncology. 2012 ; 卷 19, 編號 2. 頁 443-454.
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title = "EGFR nuclear import in gallbladder carcinoma: Nuclear phosphorylated EGFR upregulates iNOS expression and confers independent prognostic impact",
abstract = "Background: The understanding of epidermal growth factor receptor (EGFR) deregulation in carcinogenesis remains incomplete. We investigated the implications of EGFR gene status and EGFR nuclear translocation in gallbladder carcinoma (GBCA). Methods: Subcellular localization of EGFR and phosphorylated EGFR (pEGFR) was analyzed by fractional immunoblotting and confocal immunofluorescence in GBCA cell lines. pEGFR binding to iNOS promoter was assessed by chromatin immunoprecipitation with iNOS promoter activity evaluated by luciferase assay. EGFR, pEGFR, and iNOS were immunohistochemically assessable for localization and level in the training set of 104 GBCAs on tissue microarrays, with 76 cases analyzed for EGFR gene by chromogenic in situ hybridization (CISH) and mutant-enriched PCR targeting exons 19 and 21. The prognostic impact of nuclear pEGFR (N-pEGFR) immunoexpression was reaffirmed on whole sections of 58 GBCAs in the test set. Results: Nuclear expression of EGFR and pEGFR was substantiated in vitro with augmented activity of iNOS promoter elicited by pEGFR binding upon EGF treatment. Despite no mutation, EGFR amplification, identified in 11 cases (15{\%}) by CISH, strongly correlated with cytoplasmic EGFR expression (P < 0.001) but not with disease-specific survival (DSS). Immunoexpression of nuclear EGFR (N-EGFR), cytoplasmic pEGFR, and N-pEGFR was strongly related to that of iNOS (all ≤0.005). N-pEGFR independently predicted worse DSS in both training (P = 0.0468, HR = 2.024) and test sets (P = 0.0223, HR = 5.573). Conclusions: N-EGFR and N-pEGFR express in GBCA, conferring clinical aggressiveness partly through iNOS transactivation. Lacking response-predicting mutation, EGFR gene status, albeit amplified in 15{\%} of GBCA, is neither related to nuclear EGFR translocation nor prognostically useful.",
author = "Li, {Chien Feng} and Fang, {Fu Ming} and Ju-Ming Wang and Tzeng, {Ching Cherng} and Tai, {Hui Chun} and Wei, {Yu Ching} and Li, {Shau Hsuan} and Lee, {Yuan Ting} and Wang, {Yu Hui} and Yu, {Shih Chen} and Shiue, {Yow Ling} and Chu, {Patrick Yu Wei} and Wang, {Wen Ling} and Chen, {Li Tzong} and Huang, {Hsuan Ying}",
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Li, CF, Fang, FM, Wang, J-M, Tzeng, CC, Tai, HC, Wei, YC, Li, SH, Lee, YT, Wang, YH, Yu, SC, Shiue, YL, Chu, PYW, Wang, WL, Chen, LT & Huang, HY 2012, 'EGFR nuclear import in gallbladder carcinoma: Nuclear phosphorylated EGFR upregulates iNOS expression and confers independent prognostic impact', Annals of Surgical Oncology, 卷 19, 編號 2, 頁 443-454. https://doi.org/10.1245/s10434-011-1942-6

EGFR nuclear import in gallbladder carcinoma : Nuclear phosphorylated EGFR upregulates iNOS expression and confers independent prognostic impact. / Li, Chien Feng; Fang, Fu Ming; Wang, Ju-Ming; Tzeng, Ching Cherng; Tai, Hui Chun; Wei, Yu Ching; Li, Shau Hsuan; Lee, Yuan Ting; Wang, Yu Hui; Yu, Shih Chen; Shiue, Yow Ling; Chu, Patrick Yu Wei; Wang, Wen Ling; Chen, Li Tzong; Huang, Hsuan Ying.

於: Annals of Surgical Oncology, 卷 19, 編號 2, 01.02.2012, p. 443-454.

研究成果: Article

TY - JOUR

T1 - EGFR nuclear import in gallbladder carcinoma

T2 - Nuclear phosphorylated EGFR upregulates iNOS expression and confers independent prognostic impact

AU - Li, Chien Feng

AU - Fang, Fu Ming

AU - Wang, Ju-Ming

AU - Tzeng, Ching Cherng

AU - Tai, Hui Chun

AU - Wei, Yu Ching

AU - Li, Shau Hsuan

AU - Lee, Yuan Ting

AU - Wang, Yu Hui

AU - Yu, Shih Chen

AU - Shiue, Yow Ling

AU - Chu, Patrick Yu Wei

AU - Wang, Wen Ling

AU - Chen, Li Tzong

AU - Huang, Hsuan Ying

PY - 2012/2/1

Y1 - 2012/2/1

N2 - Background: The understanding of epidermal growth factor receptor (EGFR) deregulation in carcinogenesis remains incomplete. We investigated the implications of EGFR gene status and EGFR nuclear translocation in gallbladder carcinoma (GBCA). Methods: Subcellular localization of EGFR and phosphorylated EGFR (pEGFR) was analyzed by fractional immunoblotting and confocal immunofluorescence in GBCA cell lines. pEGFR binding to iNOS promoter was assessed by chromatin immunoprecipitation with iNOS promoter activity evaluated by luciferase assay. EGFR, pEGFR, and iNOS were immunohistochemically assessable for localization and level in the training set of 104 GBCAs on tissue microarrays, with 76 cases analyzed for EGFR gene by chromogenic in situ hybridization (CISH) and mutant-enriched PCR targeting exons 19 and 21. The prognostic impact of nuclear pEGFR (N-pEGFR) immunoexpression was reaffirmed on whole sections of 58 GBCAs in the test set. Results: Nuclear expression of EGFR and pEGFR was substantiated in vitro with augmented activity of iNOS promoter elicited by pEGFR binding upon EGF treatment. Despite no mutation, EGFR amplification, identified in 11 cases (15%) by CISH, strongly correlated with cytoplasmic EGFR expression (P < 0.001) but not with disease-specific survival (DSS). Immunoexpression of nuclear EGFR (N-EGFR), cytoplasmic pEGFR, and N-pEGFR was strongly related to that of iNOS (all ≤0.005). N-pEGFR independently predicted worse DSS in both training (P = 0.0468, HR = 2.024) and test sets (P = 0.0223, HR = 5.573). Conclusions: N-EGFR and N-pEGFR express in GBCA, conferring clinical aggressiveness partly through iNOS transactivation. Lacking response-predicting mutation, EGFR gene status, albeit amplified in 15% of GBCA, is neither related to nuclear EGFR translocation nor prognostically useful.

AB - Background: The understanding of epidermal growth factor receptor (EGFR) deregulation in carcinogenesis remains incomplete. We investigated the implications of EGFR gene status and EGFR nuclear translocation in gallbladder carcinoma (GBCA). Methods: Subcellular localization of EGFR and phosphorylated EGFR (pEGFR) was analyzed by fractional immunoblotting and confocal immunofluorescence in GBCA cell lines. pEGFR binding to iNOS promoter was assessed by chromatin immunoprecipitation with iNOS promoter activity evaluated by luciferase assay. EGFR, pEGFR, and iNOS were immunohistochemically assessable for localization and level in the training set of 104 GBCAs on tissue microarrays, with 76 cases analyzed for EGFR gene by chromogenic in situ hybridization (CISH) and mutant-enriched PCR targeting exons 19 and 21. The prognostic impact of nuclear pEGFR (N-pEGFR) immunoexpression was reaffirmed on whole sections of 58 GBCAs in the test set. Results: Nuclear expression of EGFR and pEGFR was substantiated in vitro with augmented activity of iNOS promoter elicited by pEGFR binding upon EGF treatment. Despite no mutation, EGFR amplification, identified in 11 cases (15%) by CISH, strongly correlated with cytoplasmic EGFR expression (P < 0.001) but not with disease-specific survival (DSS). Immunoexpression of nuclear EGFR (N-EGFR), cytoplasmic pEGFR, and N-pEGFR was strongly related to that of iNOS (all ≤0.005). N-pEGFR independently predicted worse DSS in both training (P = 0.0468, HR = 2.024) and test sets (P = 0.0223, HR = 5.573). Conclusions: N-EGFR and N-pEGFR express in GBCA, conferring clinical aggressiveness partly through iNOS transactivation. Lacking response-predicting mutation, EGFR gene status, albeit amplified in 15% of GBCA, is neither related to nuclear EGFR translocation nor prognostically useful.

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U2 - 10.1245/s10434-011-1942-6

DO - 10.1245/s10434-011-1942-6

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EP - 454

JO - Annals of Surgical Oncology

JF - Annals of Surgical Oncology

SN - 1068-9265

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ER -