Horseradish peroxidase-mediated gene therapy: choice of prodrugs in oxic and anoxic tumor conditions.
We have previously proposed the plant enzyme horseradish peroxidase (HRP) and the plant hormone indole-3-acetic acid (IAA) as an enzyme/prodrug combination for cancer gene therapy. In the current study, we evaluated the potential of HRP/IAA for gene-directed enzyme/prodrug therapy in three human tumor cell lines (T24 bladder carcinoma, MCF-7 breast adenocarcinoma, and FaDu nasopharyngeal squamous carcinoma) and one endothelial cell line (HMEC-1). The action of 10 IAA analogues in combination with HRP was studied in vitro in normoxic conditions as well as in the extreme tumor conditions of anoxia. Compounds characterized by prompt normoxic or anoxic cytotoxic activation and high HRP transfectant killing or selectivity were identified. Some variations were observed in the response of cells of different origin, with IAA, 1-Me-IAA, and 5-Br-IAA representing the most promising candidates for HRP gene therapy. In particular, 5-Br-IAA showed a very prompt and selective activation in anoxia. A strong bystander effect was produced by activated IAA and analogues because 70-90% cell kill was obtained when only 5% of the cells expressed the HRP enzyme. These results indicate that HRP/IAA represents an effective system for enzyme/prodrug-based anticancer approaches, and further improvements could be achieved by the use of novel IAA derivatives.
Greco
O
O
Gray Cancer Institute, P. O. Box 100, Northwood, Middlesex HA6 2JR, United Kingdom.
Rossiter
S
S
Kanthou
C
C
Folkes
L K
LK
Wardman
P
P
Tozer
G M
GM
Dachs
G U
GU
D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D015687:Cell Hypoxia; D003131:Combined Modality Therapy; D015316:Genetic Therapy / Q000379:methods*; D006735:Horseradish Peroxidase / Q000235:genetics*; D006801:Humans; D007210:Indoleacetic Acids / Q000494:pharmacology*; D010100:Oxygen / Q000378:metabolism; D010957:Plasmids; D011355:Prodrugs / Q000494:pharmacology*; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects* / Q000378:metabolism; D014410:Tumor Stem Cell Assay
D007210:Indoleacetic Acids; D011355:Prodrugs; D006735:Horseradish Peroxidase; D010100:Oxygen