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.
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.
Tada
Masaharu
M
Department of Gastroenterological Surgery, Kyoto University Graduate School of Medicine, 54 Kawahara-cho Shogoin Sakyo-ku Kyoto, 606-8507, Japan.
Hatano
Etsuro
E
Taura
Kojiro
K
Nitta
Takashi
T
Koizumi
Naoki
N
Ikai
Iwao
I
Shimahara
Yasuyuki
Y
D000477:Alkylating Agents; D000818:Animals; D002273:Carcinogens; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D004052:Diethylnitrosamine; D004195:Disease Models, Animal; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics; D006499:Hepatic Artery / Q000378:metabolism*; D007208:Indocyanine Green / Q000378:metabolism; D007275:Injections, Intravenous; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008297:Male; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019076:Transgenes; D001616:beta-Galactosidase / Q000235:genetics
D000477:Alkylating Agents; D002273:Carcinogens; D004052:Diethylnitrosamine; D004247:DNA; D001616:beta-Galactosidase; D007208:Indocyanine Green