EGFR nuclear import in gallbladder carcinoma: nuclear phosphorylated EGFR upregulates iNOS expression and confers independent prognostic impact

Autor: Fu Ming Fang, Yu Hui Wang, Ju Ming Wang, Yuan Ting Lee, Patrick Y. W. Chu, Yu Ching Wei, Shau Hsuan Li, Chien-Feng Li, Hui Chun Tai, Hsuan-Ying Huang, Yow-Ling Shiue, Ching Cherng Tzeng, Shih Chen Yu, Li-Tzong Chen, Wen Ling Wang
Rok vydání: 2011
Předmět:
Male
Pathology
Cytoplasm
Fluorescent Antibody Technique
Nitric Oxide Synthase Type II
medicine.disease_cause
Immunoenzyme Techniques
Tumor Cells
Cultured
Medicine
Epidermal growth factor receptor
Phosphorylation
Luciferases
Promoter Regions
Genetic
In Situ Hybridization
integumentary system
biology
Reverse Transcriptase Polymerase Chain Reaction
Prognosis
Up-Regulation
ErbB Receptors
Gene Expression Regulation
Neoplastic
Survival Rate
medicine.anatomical_structure
Oncology
Adenocarcinoma
Female
Gallbladder Neoplasms
Signal transduction
Signal Transduction
medicine.medical_specialty
Chromatin Immunoprecipitation
Blotting
Western
Active Transport
Cell Nucleus
Real-Time Polymerase Chain Reaction
Carcinoma
Humans
RNA
Messenger
Gallbladder cancer
Aged
Cell Nucleus
business.industry
medicine.disease
Carcinoma
Papillary
Cell nucleus
Mutation
Cancer research
biology.protein
Surgery
Gallbladder Neoplasm
business
Carcinogenesis
Zdroj: Annals of surgical oncology. 19(2)
ISSN: 1534-4681
Popis: 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).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.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 (P0.001) 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).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.
Databáze: OpenAIRE
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