The establishment of a Tet-on transfected human hepatoma HepG2 cell line regulated by doxycycline / 中国普通外科杂志

ABSTRACT

Objective To establish human hepatic carcinoma HepG2 cell line in which the expression of exogenous gene could be controlled by doxycycline. Methods The expression plasmid pTet-on was transfected into hepG2 cells by lipofectamine 2000.The stably transfected positive clones were screened with G 418 and subcultured. The expression level of luciferase induced by doxycycline at a concentration of 1 ?g/ml in stably transfected cells was detected one by one by transient transfection of plasmid pTRE-luc into the stably transfected cells. Results The basal expression activity of luciferase in cells of clone No 6 was 87, and the activity was 16764 after induction with doxycycline. Conclusion The HepG2/Tet-on cell line was a regulatory human hepatoma cell line, which is useful for the study of carcinogenesis and gene therapy of hepatic carcinoma.