A Preliminary Study On The Diagnostic Value Of CRP AFP Negative HBV-related HCC And CRP Biological Function

Objective:Primary liver cancer is the world’s fifth most common malignancy, and the first three causes of cancer death, there are over 50 million people worldwide suffer from liver cancer every year, of which more than half of them were in China. AFP is currently the most important and widely used serum marker for HCC. However, one-third of patients with HCC serum AFP levels were normal, which called AFP-negative HCC. Then, the diagnosis of these patients is the key issue to be addressed in HCC. Over the years, people have been looking for a new marker to assist AFP to diagnosis HCC, especially AFP-negative HCC. This paper aims to investigate the C-reactive protein (CRP) diagnostic value of AFP-negative liver cancer, as well as the biological function of CRP.Methods:Relative and absolute quantification isotope labeling technique marks the serum of 10 cases of liver cirrhosis,10 cases of AFP-negative HCC patients, and 10 cases of AFP-positive HCC patients, identified differentially expressed proteins, analyzing data to determine the target protein. By enzyme-linked immunosorbent assay (ELISA), a tissue microarray immunohistochemistry (IHC) validated screening differentially expressed proteins. The diagnostic value of the target protein were valued by further statistical analysis. In order to study the function of the target protein, we silence target protein expression of hepatoma cells HepG2, BEL-7402 to observe the ability of migration and invasion, and the effect of apoptosis and cycle. Silencing target protein expression of HepG2.2.15, the supernatant was collected to detect the titer of HBV DNA and the mRNA expression of type I interferon and downstream gene.Result:By iTRAQ technology, we screened 510 protein,14 proteins was statistically significance, including 10 down and 4 up-regulated. By Elisa, IHC, we verified the three candidate protein (CRP, C9, SAA), the result was consistent with the iTRAQ, and finally we identified the target protein CRP for further study. ROC curve analysis:CRP AFP negative HCC diagnostic sensitivity and specificity were 72.9%,60%, area under the curve of 0.724 (95% CI,0.643-0.795, p= 0.0001), CRP joint AFP area under the curve for the diagnosis of liver cancer was 0.873 (95% CI,0.829-0.910). Silence the CRP expression of HepG2, BEL-7402, the ability of invasion and migration increases, respectively, compared with the control group:47-52% and 40-49%,40-55% and 50-60%, there were no significant in apoptosis and cycle. Silencing the CRP expression of HepG2.2.15, the titer of HBV DNA was significantly higher than the control group, the mRNA expression of IFNal, IFNβ1, ISG15, OAS1, OAS2, RNA SEL was significantly increased, while the expression of OAS3 was significantly reduced.Conclusion:CRP can be used as an assist marker to diagnosis HCC, especially AFP negative HCC. CRP can inhibit the invasion and migration ability of liver cancer cells in vitro. And CRP can inhibit HBV DNA replication by type I interferon.