Proteome analysis of hepatocellular carcinoma cell strains, MHCC97‐H and MHCC97‐L, with different metastasis potentials

Abstract To better understand the mechanism underlying hepatocellular carcinoma (HCC) metastasis and to search for potential markers for HCC prognosis, differential proteome analysis on two HCC cell strains with high and low metastatic potentials, MHCC97‐H and MHCC97‐L, was conducted using two‐dimensional (2‐D) gel electrophoresis followed by matrix‐assisted laser desorption/time of flight mass spectrometry and liquid chromatography ion trap mass spectrometry. Image analysis of silver‐stained 2‐D gels revealed that 56 protein spots showed significant differential expression in MHCC97‐H and MHCC97‐L cells (Student's t ‐test, P < 0.05) and 4 protein spots were only detected in MHCC97‐H cells. Fourteen protein spots were further identified using in‐gel tryptic digestion, peptide mass fingerprinting and tandem mass spectrometry. The expressions of pyruvate kinase M2, ubiquitin carboxy‐terminal hydrolase L1, laminin receptor 67 kDa, S100 calcium‐binding protein A4, thioredoxin and cytokeratin 19 were elevated in MHCC97‐H cells. However, manganese superoxide dismutase, calreticulin precursor, cathepsin D, lactate dehydrogenase B, non‐metastatic cell protein 1, cofilin 1 and calumenin precursor were down‐regulated in MHCC97‐H cells. Intriguingly, most of these identified proteins have been reported to be associated with tumor metastasis. The functional implications of alterations in the levels of these proteins are discussed.