title: A multiplex tissue immunoblotting assay for proteomic profiling: a pilot study of the normal to tumor transition of esophageal squamous cell carcinoma.
abstract: Esophageal cancer remains a highly lethal malignancy for which the genetic and proteomic events are poorly understood. Studies have reported dysregulated proteins in esophageal carcinoma; however, the magnitude of these changes remains largely uncharacterized. Little is known about alterations early in the neoplastic pathway. Using multiplex tissue immunoblotting, we quantified the expression of seven proteins in esophageal carcinogenesis. Regions of normal, dysplasia, and invasive carcinoma of the squamous esophagus in six patients were characterized. Pan-cytokeratin (CK) was essentially unchanged across the transition (0.96 in dysplasia and 0.69 in tumor). Expression levels of annexin 1, CK-4, and CK-14 were all decreased in dysplasia and tumor compared with normal (reference, 1.00): annexin 1, 0.30 in dysplasia and 0.15 in tumor; CK-4, 0.20 in dysplasia and 0.16 in tumor; and CK-14, 0.54 in dysplasia and 0.40 in tumor. Expression of two proteins was increased in dysplasia and tumor versus normal: cyclooxygenase-2, 1.35 in dysplasia and 2.32 in tumor and p53, 1.29 in dysplasia and 2.37 in tumor. Secreted protein, acidic and rich in cysteine, which is expressed in the adjacent stroma, was 1.56-fold higher in stroma underlying dysplasia and 6.20-fold increased in dysplastic stroma surrounding invasive tumor. These findings suggest that changes in protein expression can be detected during the transition to dysplasia and may be useful biomarkers.
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland, USA.
authors.forename: Joon-Yong
authors.initials: JY
authors.lastname: Chung
authors.forename: Till
authors.initials: T
authors.lastname: Braunschweig
authors.forename: Nan
authors.initials: N
authors.lastname: Hu
authors.forename: Mark
authors.initials: M
authors.lastname: Roth
authors.forename: June L
authors.initials: JL
authors.lastname: Traicoff
authors.forename: Quan-Hong
authors.initials: QH
authors.lastname: Wang
authors.forename: Vladimir
authors.initials: V
authors.lastname: Knezevic
authors.forename: Philip R
authors.initials: PR
authors.lastname: Taylor
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Hewitt
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D018450:Disease Progression; D004938:Esophageal Neoplasms / Q000378:metabolism* / Q000473:pathology; D004947:Esophagus / Q000378:metabolism* / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D015151:Immunoblotting; D007633:Keratins / Q000378:metabolism; D025202:Molecular Diagnostic Techniques; D009361:Neoplasm Invasiveness / Q000175:diagnosis; D015676:Osteonectin / Q000378:metabolism; D010865:Pilot Projects; D011230:Precancerous Conditions / Q000378:metabolism* / Q000473:pathology; D020543:Proteome / Q000032:analysis*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D015676:Osteonectin; D020543:Proteome; D016159:Tumor Suppressor Protein p53; D007633:Keratins