[Two-dimensional electrophoresis analysis of hydrophobic membrane proteome in mouse hepatocarcinoma cell H22]

To compare the membrane protein profile of mouse hepatocarcinoma cell H22 with that of normal liver cell.

The membrane proteins in mouse hepatocarcinoma cell H22 and normal liver cell were extracted and their concentrations were determined by Bradford method. The proteins were separated by two-dimensional electrophoresis, and then stained with silver. The 2-DE maps were scanned and analyzed by Image Master 2D Platinum software. The differential expression protein spots were cut out from the gels, and the peptide fingerprinting was determined by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectrometry), followed by matching to Swiss-Prot protein database by Aldente software with experimental pI and MW data.

Compared to normal liver cells, 8 membrane proteins, including sulfatase-modifying factor 2, protein kinase C and casein kinase II substrate protein 3, sorting and assembly machinery component 50 homolog, macrophage scavenger receptor types I/II, uncharacterized protein C9 or f135 homolog, tight junction protein ZO-2, 3-hydroxy-3- methylglutaryl-coenzyme A reductase, and vacuolar protein sorting-associated protein 52 homolog were upregulated in H22 cells.

The membrane proteins involved in cell metabolism, proliferation, signal transduction, and skeleton, which are highly expressed in mouse hepatocarcinoma H22 cells, are probably related to the proliferation, invasion and migration of this tumor cell line.