Glypican-3 (GPC3) is overexpressed in hepatocellular carcinoma (HCC) but not in chronic hepatitis (CH) and liver cirrhosis (LC). We have reported the possibility of GPC3-specific cytotoxic T lymphocytes (CTLs) serving as a marker for the early diagnosis of imaging invisible HCC. In this study, to identify new early diagnostic biomarker of imaging invisible HCC, we analyzed plasma of healthy donors and patients with CH, LC and HCC using surface-enhanced laser desorption-ionaization time-of-flight mass spectrometry (SELDI-TOF-MS). The intensities of four peaks were significantly increased in HCC patients compared with healthy donors. Two of these four peaks were significantly higher in CH and LC patients with GPC3-specific CTLs than in those without. One peak (11.7 kDa) was predicted to be β2-microglobulin (β2-MG) by molecular mass. There was a correlation between concentration of β2-MG by latex agglutination immunoassay in plasma and peak intensity using SELDI-TOF-MS. The 11.7 kDa protein was fractionated by gel filtration and was identified as β2-MG by Western blot analysis. These results suggest that the level of β2-MG in plasma from patients with CH and LC could be a useful marker for the early diagnosis of imaging invisible HCC, however further investigation is needed.

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