Expression difference and function research of family with sequence similarity 96, member B in human hepatocellular carcinoma

Abstract

Objective To study the expression difference of family with sequence similarity 96,member B (FAM96B) in human hepatocellular carcinoma (HCC) and its impact on the proliferation and apoptosis of HepG2 cell line.Methods FAM96B mRNA and protein expression was examined respectively by real-time fluorogenic quantitative polymerase chain reacton (FQ-PCR) and Western blotting in tumor tissues and adjacent tissues from 42 patients with HCC.The FAM96B mRNA and protein expression was detected in HepG2 and L-02 cells.Moreover the eukaryotic vector of human FAM96B was constructed and transfected into HepG2 cells.The proliferative rate and apoptosis rate of HepG2 cells were examined by the methyl thiazol tetrazolium (MTT) assay and fluorescence-activated cell sorting (FACS).Results In 42 patients with HCC,the global mRNA expression of FAM96B was 0.304 ± 0.094 times in tumor tissues than in adjacent tissues.And its mRNA expression was 0.406 ±0.115 times in HepG2 cell than in L-02 cell.While the Western blotting analysis confirmed that FAM96B protein expression in tumor tissues was 0.288±0.132 times lower than in the adjacent tissues (P ＜ 0.05).And FAM96B protein expression in HepG2 cell was 0.359 ± 0.143 times lower than in L-02 cell.The MTT assay showed that the A value inFAM96B group was lower than in control group,and overexpression of FAM96B significantly inhibited proliferation of HepG2 cells (P ＜0.05).FACS analysis indicated that there was significant difference in cell apoptosis rate between FAM96B-expressing clones group and the control group (vector-transfected cells group) (P ＜ 0.05).Conclusion FAM96B expression showed statistically significant difference between tumor tissues and adjacent tissues in 42 cases of HCC.Moreover FAM96B expression in HepG2 cells was significantly lower than in L-02 cells (P ＜ 0.05).Furthermore,overexpression of FAM96B can induce apoptosis of HepG2 cells and inhibit proliferation of HepG2 cells.The data suggest that FAM96B probably plays an important role in carcinogenesis of HCC. Key words: Carcinoma, hepatocellular; Family with sequence similarity 96, member B; Proliferation