title: Separation and identification of differentially expressed nuclear matrix proteins between human esophageal immortalized and carcinomatous cell lines.
abstract: OBJECTIVE
  To separate and identify differentially expressed nuclear matrix proteins (NMPs) between the immortalized human esophageal epithelial cell line (SHEE) and the malignantly transformed esophageal carcinoma cell line (SHEEC), and to provide new ways for finding specific markers and the pathogenesis of esophageal carcinoma.
  
  
  METHODS
  SHEE and SHEEC cell lines were used to extract NMPs. The quality of NMPs was monitored by Western blot analysis including DNA topoisomerase IIalpha, proliferation cell nuclear antigen (PCNA) and histone. NMPs of SHEE and SHEEC were analyzed by two-dimensional electrophoresis (2-DE), silver staining and PDQuest6.2 image analysis software. Three spots in which the differentially expressed NMPs were more obvious, were selected and analyzed with matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI- TOF-MS) and database search.
  
  
  RESULTS
  Western blot analysis revealed that DNA topoisomerase IIalpha and PCNA were detected, and the majority of histones were deleted in NMPs of SHEE and SHEEC. After 2-DE image analysis by PDQuest6.2 software, the 2-DE maps were detected with an average of 106+/-7.1 spots in SHEE and 132+/-5.0 spots in SHEEC. Most of them were matched one another (r=0.72), only 16 protein spots were found differing in intensity. Three NMPs including cytoskeletal tropomyosin, FK506-binding protein 6, similar to retinoblastoma binding protein 8 were preliminarily identified by MALDI- TOF-MS.
  
  
  CONCLUSIONS
  These differentially expressed NMPs may play an important role during malignant transformation from SHEE to SHEEC. Their separation and identification will contribute to searching for specific markers and probing into the pathogenesis of esophageal carcinoma.
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Shantou University, 22 Xinling Road, Shantou 515031, Guangdong Province, China.
authors.forename: Xing-Dong
authors.initials: XD
authors.lastname: Xiong
authors.forename: En-Min
authors.initials: EM
authors.lastname: Li
authors.forename: Li-Yan
authors.initials: LY
authors.lastname: Xu
authors.forename: Hai-Bin
authors.initials: HB
authors.lastname: Chen
authors.forename: Ling
authors.initials: L
authors.lastname: Chen
authors.forename: Wei-Jia
authors.initials: WJ
authors.lastname: Cai
authors.forename: Ya-Li
authors.initials: YL
authors.lastname: Han
authors.forename: Zhong-Ying
authors.initials: ZY
authors.lastname: Shen
authors.forename: Yi
authors.initials: Y
authors.lastname: Zeng
mesh_terms: D015153:Blotting, Western; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D004938:Esophageal Neoplasms; D004947:Esophagus / Q000737:chemistry* / Q000166:cytology*; D006801:Humans; D034521:Nuclear Matrix-Associated Proteins / Q000032:analysis* / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D034521:Nuclear Matrix-Associated Proteins