Construction of eukaryotic expression vector carrying human TSLC1 gene and its expression in HepG2 cells

Objective:To construct the eukaryotic expression vector for human TSLC1 gene,and to express TSLC1 in HepG2 cells for investigating its effect on HepG2 cell growth.Methods:Full length of TSLC1 cDNA was amplified from RNA of normal human liver by RT-PCR,and cloned into pCI-neo expression vector.The recombinant plasmid pCI-TSLC1 was identified with restriction enzyme and sequenced,and then was stably transfected into HepG2 cells with lipofectamine 2000. The positive clones were examined by western-blotting and immunofluorescence,cell growth was analyzed with MTT assay. Results:The eukaryotic expression vector pCI-TSLC1 was successfully constructed and the stable cell line highly expressing TSLC1 protein was obtained.The growth of TSLC1-transfected cells was significantly suppressed in vitro.Conclusion:The HepG2 stable cell line could highly express TSLC1 protein,which provided a basis for further exploring the roles of TSLC1 in hepatocellular carcinoma.