Cre/loxP system controlled by specific promoter for radiation-mediated gene therapy of hepatoma

Anticancer Res. 2007 May-Jun;27(3B):1571-9.

Abstract

Background: To specifically target malignant cells, cancer gene therapy needs to combine highly selective gene delivery with highly specific gene expression. In this study, hepatitis B virus (HBV) enhancer II was combined with alpha-fetoprotein (AFP) promoter which selectively controls the Cre/loxP system in hepatoma.

Materials and methods: pE4luc, pE4Tk, EIIAPA-Cre and E4CMV-STOP-Tk constructs and AFP promoter combined with HBV enhancer were constructed and transfected into HepG2, HeLa and NIH-3T3 cell lines.

Results: The E4 enhancer showed the highest luciferase gene expression at a dose range of 5 approximately 7 Gy after 60 hours irradiation. The EIIAPA chimeric promoter which controls the Cre/loxP system provided high specificity only to the hepatoma cells. In addition, the E4 response to radiation encoded more Herpes simplex virus thymidine kinase (HSV1-Tk) protein and killed more tumor cells.

Conclusion: The chimeric EIIAPA promoter can precisely control the Cre/loxP switch and the radiation effect on the EIIAPA-Cre and E4CMV-STOP-Tk system shows promising results in terms of cell survival of hepatocellular carcinoma (HCC).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology
  • Carcinoma, Hepatocellular / therapy*
  • Cell Survival
  • Enhancer Elements, Genetic / radiation effects*
  • Ganciclovir / pharmacology
  • Genetic Therapy / methods*
  • HeLa Cells
  • Hepatitis B virus / genetics
  • Herpesvirus 1, Human / genetics
  • Humans
  • Integrases / genetics*
  • Liver Neoplasms / therapy*
  • Mice
  • Mice, SCID
  • NIH 3T3 Cells
  • Promoter Regions, Genetic
  • Radiation
  • Thymidine Kinase / genetics
  • Transfection
  • Xenograft Model Antitumor Assays
  • alpha-Fetoproteins / genetics

Substances

  • Antiviral Agents
  • alpha-Fetoproteins
  • Thymidine Kinase
  • Cre recombinase
  • Integrases
  • Ganciclovir