High volume hydrodynamic injection of plasmid DNA via the hepatic artery results in a high level of gene expression in rat hepatocellular carcinoma induced by diethylnitrosamine.

Abstract

BACKGROUND:

Hydrodynamic injection of naked plasmid DNA (pDNA) via the tail vein is a safe and effective method of gene transfer to the liver. However, successful gene transfer has yet to be shown for hepatocellular carcinoma (HCC); therefore, we investigated the feasibility and efficacy of hydrodynamic injection via the tail vein and hepatic artery in a diethylnitrosamine (DEN)-induced HCC model in rats.

METHODS:

HCC was induced in Sprague-Dawley rats by 100 ppm DEN in drinking water. pCMV-SPORT-beta-galactosidase (beta-gal, 400 microg) was injected (i) via the tail vein in a volume of 0.1 ml/g in 30 s or (ii) via the hepatic artery in a volume of 5 or 10 ml at 1 ml/s, either with or without temporary occlusion of the inferior vena cava (IVC) and portal vein (PV). The liver was harvested 24 h after administration, and beta-gal expression was evaluated with X-gal staining and measurement of enzymatic activity in tissue homogenates.

RESULTS:

Hydrodynamic injection via the tail vein achieved transgene expression only in non-cancerous tissue (tumor: 0.16 +/- 0.04%, non-tumor: 5.07 +/- 1.66%). Hydrodynamic injection via the hepatic artery was tolerated, but failed to produce efficient transgene expression in tumor and non-tumor cells. On the other hand, concomitant use of temporary IVC/PV occlusion with hydrodynamic injection via the hepatic artery dramatically increased transgene expression in cancer cells, but tumor-selective gene transfer was not achieved with this procedure (tumor: 7.38 +/- 3.66%, non-tumor: 7.77 +/- 1.06%).

CONCLUSIONS:

High-volume hydrodynamic injection of a pDNA solution via the hepatic artery with IVC/PV occlusion achieved a high level of gene expression in a HCC rat model. This gene transfer technique may have potential in clinical gene therapy for HCC.

• Authors: Masaharu Tada, Etsuro Hatano, Kojiro Taura, Takashi Nitta, Naoki Koizumi, Iwao Ikai, Yasuyuki Shimahara
• Journal: The journal of gene medicine (J Gene Med) Vol. 8 Issue 8 Pg. 1018-26 (Aug 2006) ISSN: 1099-498X [Print] England
• PMID: 16779866 (Publication Type: Comparative Study, Journal Article)
• Copyright: 2006 John Wiley & Sons, Ltd.

Chemical References

• Alkylating Agents
• Carcinogens
• Diethylnitrosamine
• DNA
• beta-Galactosidase
• Indocyanine Green

Topics

• Alkylating Agents
• Animals
• Carcinogens
• Carcinoma, Hepatocellular (metabolism, pathology, therapy)
• Cytomegalovirus (genetics)
• DNA (administration & dosage, genetics)
• Diethylnitrosamine
• Disease Models, Animal
• Gene Expression
• Gene Transfer Techniques
• Genetic Therapy (methods)
• Genetic Vectors (chemistry, genetics)
• Hepatic Artery (metabolism)
• Indocyanine Green (metabolism)
• Injections, Intravenous
• Liver Neoplasms (metabolism, pathology, therapy)
• Male
• Plasmids (administration & dosage, genetics)
• Rats
• Rats, Sprague-Dawley
• Transgenes
• beta-Galactosidase (genetics)