Study on expression of Beta-galactoside alpha-2,6-sialyltransferase Ⅰand HCC specific carbohydrate chain of alpha fetoprotein in hepatocellular carcinoma cell line and liver disease blood
Hepatocellular carcinoma (HCC) is an important malignant tumor, which is high morbidity and poor prognosis. It is important for the clinical diagnosis and treatment of HCC to discover and distinguish HCC in liver diseases precisely as early as possible. In the early stage of tumor genesis, it is hard to discover HCC because of the limitation of instrument and the deep location of liver, so serological biomarker is needed for HCC early diagnosis.
AFP as a serological biomarker is used widely. But the sensitivity and specificity of AFP is not high enough, there are lots of studies in progress to find more effective methods for HCC diagnosis. Beta-galactoside alpha-2,6-sialyltransferase Ⅰ(ST6GalⅠ), regulating the sialoglycans of proteins, is one member of the sialytransgerases family. Some studies showed that ST6GalⅠ was increasingly expressed in HCC, and may be released into blood with the increasing malignance of hepatocytes. ST Gal Ⅰmay be a potential marker for HCC early diagnosis. Otherwise, it was reported than the carbohydrate chains of HCC was different from liver cirrhosis, so, the specific carbohydrate chains of AFP in HCC may be the potential biomarker for HCC early diagnosis.
Objective：In order to find biomarker for HCC early diagnosis, ST6Gal Ⅰand the carbohydrate chain of AFP were studied in this research.
Methods: 1) ST6GalⅠpolyclonal antibody was prepared by immuning rabbit with synthesized peptide of ST6GalⅠ; 2) Indirect ELISA, western blot and immunocytochemistry were used to study the antiserum; 3) Western blot was used to study the expression of ST6GalⅠin liver disease blood; 4) Immunoprecipitation was used to detect TF,H2,Lewis Y in HCC cell line and blood.
Results: 1) The titer of antiserum was tested to be 1:400 000 by indirect ELISA, the antiserum could detect ST6GalⅠin HCC cell line by western blot and immunocytochemistry; 2) ST6GalⅠexpressed in HCC plasma (3/3 cases) and HCC serum (2/2 cases) without fail, in liver cirrhosis plasma (1/2 cases) and liver cirrhosis serum (1/2cases) and normal plasma (2/3cases) partially; 3) TF, H2 and Lewis Y were positive in HepG2 and HCC plasma by immunoprecipitation.
Conclusion: ST6GalⅠ and the specific carbohydrate chain of AFP were studied. A kind of rabbit anti-human ST6GalⅠ polyclonal antibody was prepared to detect the ST in HCC cell line and blood by immunology method. And the polyclonal antibody could be used for the research of ST6GalⅠ. ST6GalⅠhas different expression in HCC and liver cirrhosis, it may be the potential biomarker for HCC early diagnosis. TF, H2 and Lewis Y are the specific carbohydrate chain of AFP in HCC, these carbohydrate chains was detected in AFP of HCC cell line and HCC plasma by Immunoprecipitation. The specific carbohydrate chain of AFP in HCC may be a joint biomarker for improving the sensitivity and specificity of AFP in HCC diagnosis.
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