title: 2D differential in-gel electrophoresis for the identification of esophageal scans cell cancer-specific protein markers.
abstract: The reproducibility of conventional two-dimensional (2D) gel electrophoresis can be improved using differential in-gel electrophoresis (DIGE), a new emerging technology for proteomic analysis. In DIGE, two pools of proteins are labeled with 1-(5-carboxypentyl)-1'-propylindocarbocyanine halide (Cy3) N-hydroxy-succinimidyl ester and 1-(5-carboxypentyl)-1'-methylindodi-carbocyanine halide (Cy5) N-hydroxysuccinimidyl ester fluorescent dyes, respectively. The labeled proteins are mixed and separated in the same 2D gel. 2D DIGE was applied to quantify the differences in protein expression between laser capture microdissection-procured esophageal carcinoma cells and normal epithelial cells and to define cancer-specific and normal-specific protein markers. Analysis of the 2D images from protein lysates of approximately 250,000 cancer cells and normal cells identified 1038 protein spots in cancer cell lysates and 1088 protein spots in normal cell lysates. Of the detected proteins, 58 spots were up-regulated by >3-fold and 107 were down-regulated by >3-fold in cancer cells. In addition to previously identified down-regulated protein annexin I, tumor rejection antigen (gp96) was found up-regulated in esophageal squamous cell cancer. Global quantification of protein expression between laser capture-microdissected patient-matched cancer cells and normal cells using 2D DIGE in combination with mass spectrometry is a powerful tool for the molecular characterization of cancer progression and identification of cancer-specific protein markers.
authors.affiliation: Department of Biochemistry University of Texas Southwestern Medical Center, Dallas, Texas 75390-9038, USA.
authors.forename: Ge
authors.initials: G
authors.lastname: Zhou
authors.forename: Hongmei
authors.initials: H
authors.lastname: Li
authors.forename: Dianne
authors.initials: D
authors.lastname: DeCamp
authors.forename: She
authors.initials: S
authors.lastname: Chen
authors.forename: Hongjun
authors.initials: H
authors.lastname: Shu
authors.forename: Yi
authors.initials: Y
authors.lastname: Gong
authors.forename: Michael
authors.initials: M
authors.lastname: Flaig
authors.forename: John W
authors.initials: JW
authors.lastname: Gillespie
authors.forename: Nan
authors.initials: N
authors.lastname: Hu
authors.forename: Philip R
authors.initials: PR
authors.lastname: Taylor
authors.forename: Michael R
authors.initials: MR
authors.lastname: Emmert-Buck
authors.forename: Lance A
authors.initials: LA
authors.lastname: Liotta
authors.forename: Emanuel F
authors.initials: EF
authors.lastname: Petricoin
authors.forename: Yingming
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics* / Q000302:isolation & purification*; D002294:Carcinoma, Squamous Cell / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004938:Esophageal Neoplasms / Q000235:genetics*; D020869:Gene Expression Profiling; D013058:Mass Spectrometry; D009363:Neoplasm Proteins / Q000235:genetics* / Q000302:isolation & purification*; D020543:Proteome / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D020543:Proteome