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. 2002 Oct;8(5):777-81.
doi: 10.3748/wjg.v8.i5.777.

Identification of differentially expressed proteins between human esophageal immortalized and carcinomatous cell lines by two-dimensional electrophoresis and MALDI-TOF-mass spectrometry

Xing-Dong Xiong  1 , Li-Yan Xu, Zhong-Ying Shen, Wei-Jia Cai, Jian-Min Luo, Ya-Li Han, En-Min Li
Affiliations

Identification of differentially expressed proteins between human esophageal immortalized and carcinomatous cell lines by two-dimensional electrophoresis and MALDI-TOF-mass spectrometry

Xing-Dong Xiong et al. World J Gastroenterol. 2002 Oct.

Abstract

Aim: To identify the differentially expressed proteins between the human immortalized esophageal epithelial cell line (SHEE) and the malignant transformed esophageal carcinoma cell line (SHEEC), and to explore new ways for studying esophageal carcinoma associated genes.

Methods: SHEE and SHEEC cell lines were used to separate differentially expressed proteins by two-dimensional electrophoresis. The silver-stained 2-D gels was scanned with EDAS290 digital camera system and analyzed with the PDQuest 6.2 Software. Six spots in which the differentially expressed protein was more obvious were selected and analyzed with matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).

Results: There were 107+/-4.58 and 115+/-9.91 protein spots observed in SHEE and SHEEC respectively, and the majority of these spots between the two cell lines matched each other (r=0.772), only a few were expressed differentially. After analyzed by MALDI-TOF-MS and database search for the six differentially expressed proteins, One new protein as well as other five sequence-known proteins including RNPEP-like protein, human rRNA gene upstream sequence binding transcription factor, uracil DNA glycosylase, Annexin A2 and p300/CBP-associated factor were preliminarily identified.

Conclusion: These differentially expressed proteins might play an importance role during malignant transformation of SHEEC from SHEE. The identification of these proteins may serve as a new way for studying esophageal carcinoma associated genes.

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Figures

Figure 1
Figure 1
The differentially expressed protein spots observed in SHEE (left) and SHEEC (right) two-dimensional gels samples.
Figure 2
Figure 2
The MALDI-TOF mass spectrum map of protein spot 1. Mode of operation: Reflector Extraction mode: delayed Accelerating voltage: 20000 V. Acquisition mass range: 750-3500 Da Number of laser shots: 150/spectrum Laser intensity: 2233
Figure 3
Figure 3
The MALDI-TOF mass spectrum map of protein spot 6. Mode of operation: Reflector Extraction mode: delayed Accelerating voltage: 20000 V. Acquisition mass range: 750-3000 Da Number of laser shots: 200/spectrum Laser intensity: 2224
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