Humoral autoimmune response to nucleophosmin in the immunodiagnosis of hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a worldwide prevalent cancer with an exremely poor prognosis. Detection of serum α-fetoprotein (AFP) and liver imaging techniques are the conventional methods used clinically for the identification of this malignancy. However, these techniques are not reliable for early diagnosis, and particularly the sensitivity and specificity of AFP in HCC diagnosis are not optimal. Therefore, there is an urgent need for the development of more sensitive and specific methods that can improve AFP quantification in the early detection of HCC. In the present study, autoantibody responses to nucleophosmin (NPM1) in HCC patients were evaluated by enzyme-linked immunosorbent assay (ELISA), western blotting and indirect immunofluorescence. Immunohistochemistry (IHC) with tissue array slides was also performed to analyze protein expression of NPM1 in HCC and control tissues. The prevalence of autoantibodies against NPM1 was 22.4% (17/76) in HCC, which was significantly higher than that in sera from patients with liver cirrhosis (LC), chronic hepatitis (CH) and systemic lupus erythematosus (SLE) (P<0.01). The average titer of autoantibodies against NPM1 in HCC sera was higher compared to that in LC, CH, SLE and normal human sera (NHS) (P<0.01). In addition, anti-NMP1 autoantibodies were detected in sera from several HCC patients with serial bleeding samples. A stronger reactive band corresponding to NMP1 was visualized in the western blot analyses, utilizing sera from patients 3-6 months before the clinical diagnosis of HCC. Our data indicate that NPM1 and the anti-NPM1 system may have potential as an early-stage biomarker for HCC screening and diagnosis.