Preparation Antibody And Immunohistochemical Study Of Human Hepatocellular Carcinoma-associated Antige SMP-30

OBJECTIVE: Expressed MBP-SMP-30 fusion protein and that was used to immunize rabbit to prepare the anti-MBP-SMP-30 polyclonal antibody. Immunohistochemical staining was used to detecte the expression of SMP-30 protein in HCC and study its possible role and clinical significance in tumorigenesis and development.METHODS: (1)The cDNA that represents 165 amino acids of SMP-30 in its C-end was recombined in expressing plasmid.The MBP-SMP-30 fusion protein was expressed in E.coli and was purified by affinity chromatographer. The MBP-SMP-30 fusion protein was used to immunize rabbit to prepare the anti-MBP-SMP-30 polyclonal antibody. The polyclonal antibody was identified by ELISA and Western blot(.2)One normal kidney and six normal liver samples, 48 cases of HCC and their corresponding paracancerous tissues were used for detection. Immunohistochemical staining for SMP-30 protein was performed with SABC method on paraffin sections. The results associated with clinical data were analyzed.RESULTS: (1)The polyclonal antibody was found to specifically bind to MBP-SMP-30 in ELISA and Western blot analysis(.2)SMP-30 was found in the cytoplasm and nucleus of hepatocyte and renal proximal tubular epithelium. (3) The protein expression level in paracancerous tissues was higher than that in HCC(P<0.05).(4) The expression of SMP-30 protein was no correlation with sex, age, tumor size, pathological grade, hepatic cirrhosis, HBsAg and AFP...