Two-dimensional gel electrophoresis analysis of the proteomes expressed in the human hepatoma cell line BEL-7404 and normal liver cell line L-02 |
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Authors: | Lirong Yu Nan Wang Gaode Wu Yonghua Xu Qichang Xia |
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Institution: | (1) Shanghai Institute of Biochemistry, Chinese Academy of Sciences, 200031 Shanghai, China;(2) Shanghai Institute of Cell Biology, Chinese Academy of Sciences, 200031 Shanghai, China |
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Abstract: | Proteome analysis technology has been used extensively in conducting discovery research of biology and has become one of the
most essential technologies in functional genomics. The proteomes of the human hepatoma cell line BEL-7404 and the normal
human liver cell line L-02 have been separated by high resolution two-dimensional gel electrophoresis (2-DE) with immobilized
pH gradient isoelectric focusing (IPG-IEF) in the first dimension and sodium dodecyl sulfate polyacrylamide gel electrophoresis
(SDS-PAGE) in the second dimension (IPG-DALT). The resulting images have been analyzed using 2-D analysis software. Quantitative
analysis reveals that 7 protein spots are detected only in hepatoma BEL-7404 cells, 14 only in L-02 cells, and 78 protein
spots show significant fluctuation in quantity in both cell lines (P< 0.01). These protein spots have been displayed on a proteome differential expression map. Analysis for the reproducibility
of 2-DE indicates that the positional variability in the IEF dimension is 0.73 mm, while the variability in the SDS-PAGE dimension
is 0.44 mm, and the quantitative variability is 17.6%–19.2%. These results suggest that the reproducibility of 2-DE has been
suitable for the study of differential expression of proteomes. Proteome differential expression maps can be useful tools
for disease diagnosis, drug-target validation analysis and biological process elucidation. |
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Keywords: | proteome two-dimensional gel electrophoresis differential expression map human hepatoma cells liver cells |
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