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Establishment of a Doxycycline-regulated Cell Line with Inducible, Doubly-stable Expression of the Wild-type P53 Gene from P53-deleted Hepatocellular Carcinoma Cells by Tian-Yi Chi; George G Chen; Lok-Kee Ho; Paul BS Lai is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Establishment of a Doxycycline-regulated Cell Line with Inducible, Doubly-stable Expression of the Wild-type P53 Gene from P53-deleted Hepatocellular Carcinoma Cells about?

p53 is important in the development of hepatocellular carcinoma (HCC) and in therapeutic approaches, but the mechanism whereby it inhibits HCC growth is still unclear. The aim of the present study was to establish a HCC cell system in which p53 levels can be regulated. Full-length wild-type p53 cDNA obtained by PCR was cloned into a retroviral response vector controlled by the tetracycline responsive element (RevTRE-p53). The regulatory vectors RevTet-Off and RevTRE-p53 were transfected into a packaging cell line, PT67. Hep3B cells in which the p53 gene was deleted were infected with RevTet-Off viral particles from the PT67. Three G418-resistant cell clones with high luciferase expression and low background were infected with RevTRE-p53. By screening dozens of RevTRE-p53-infected clones with hygromycin we identified the one with the highest expression of p53 and the lowest background after doxycycline treatment. The results showed that p53 expression in this cell clone could be simply turned on or off by removing or adding doxycycline. Furthermore, it was found that the level of p53 protein was negatively and sensitively related to the doxycycline concentration. In conclusion, we h

Who reads Establishment of a Doxycycline-regulated Cell Line with Inducible, Doubly-stable Expression of the Wild-type P53 Gene from P53-deleted Hepatocellular Carcinoma Cells?

Author

Tian-Yi Chi; George G Chen; Lok-Kee Ho; Paul BS Lai

Publisher

BioMed Central; Springer (Biomed Central Ltd.); London: BioMed Central, 2001-; Springer Science and Business Media LLC (ISSN 1475-2867)

Published

2005

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)