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Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, particularly prevalent in Asia, with very poor prognosis. Although α-fetoprotein (AFP) is the most effective biomarker available to detect HCC, it has a low clinical sensitivity and specificity as biomarker. Therefore, there is a need for the development of more sensitive and specific assays that can supplement AFP for the early detection of liver cancer. Recently, autoantibodies, which are generated by immune system recognizing the presence of the abnormal tumor-associated antigens, are suggested as promising biomarkers for the early detection of tumors. In this study, to improve the early detection and diagnosis of HCC, determination of autoantibody responses to AFP in HCC patients was performed and analyzed its correlation with the progression of disease. The serum AFP levels were measured using sandwich ELSA commercially available and anti-AFP autoantibody levels were determined by ELISA using purified AFP from HepG2 cells as coating antigen. Assays were performed for 50 healthy controls, 50 patients with HCC, 36 patients with liver cirrhosis, and 35 patients with chronic hepatitis. Anti-AFP autoantibodies were detected in patients’ sera showing elevated AFP levels, which revealed that AFP secreted from tumor cells elicit immune response which induce tumor-associated autoantibodies. Even when the AFP level was very low anti-AFP responses were detectable, impling that autoantibody response to AFP can be used as an early detection biomarker. Several patient sera showing highly elevated AFP, however, did not show anti-AFP antibody response, which might be related to the intrinsic property of the AFP molecule as an immune suppressor. In addition, simultaneous detection of AFP and anti-AFP response in patients’ sera improved the diagnosis of HCC and we discuss about its usage as HCC diagnosis biomarker.