摘要
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.
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 TSLCl-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.
