Establishment of HepG2~(Tet-on) cell line controlled by the Tet-on regulatory system / 中国普通外科杂志

ABSTRACT

Objective To establish tetracycline-controlled inducible system(Tet-on) in HepG2 cell for further research of the function of related gens.Methods The HepG2 cells were transfected with pWHE146 vector by using liposome transfection reagent.The transfected cells were selected in medium containing G418,and G418-resistant clones were isolated.All individual G418-resistant clones were screened by transient transfection with plasmid pTRE-hyg-luc for clones with low background and high induction of luciferase in response to Dox.Results One HepG2 cell line,which exhibited high levels of induction(154,106 times)and high gene expression levels,was obtained.Conclusions The HepG2 cell line can be used to highly express eukaryotic gene and this Tet-on system is available for use in eukaryotic gene function studies.