Gene therapy for alpha-fetoprotein-producing human hepatoma cells by adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene.
We have developed a recombinant replication-defective adenovirus containing human alpha-fetoprotein (AFP) promoter/enhancer to direct cell type-specific expression of the herpes simplex virus thymidine kinase (HSVtk) gene to AFP-producing hepatocellular carcinoma (HCC) cells. After an in vitro infection by a recombinant adenovirus carrying the lacZ gene under the control of human AFP promoter/enhancer (AdAFPlacZ), an expression of the lacZ gene was demonstrated efficiently in AFP-producing HuH-7 and HepG2 cell lines, but not in AFP-nonproducing HLE and HLF cell lines, although lacZ gene expression was demonstrated in all these cell lines when infected with adenovirus vector carrying lacZ gene driven by the beta-actin-based promoter. Expression of the HSVtk gene by adenovirus, from AFP promoter/enhancer (AdAFPtk) induced the cells sensitive to ganciclovir (GCV) in the AFP-producing cell line efficiently, but not in AFP-nonproducing HLF hepatoma cells. An in vitro bystander effect was observed when only 10% of the cells were infected with AdAFPtk. These findings suggest that the AFP promoter/enhancer sequence can provide the tumor-specific activity for the therapeutic gene expression, and that the AdAFPtk vector induces the selective growth inhibition by GCV in the adenovirus-infected human hepatoma cells in vitro. Recombinant adenovirus transfer of the HSVtk gene under the control of tumor-specific promoter followed by GCV may have promise as a targeted in situ treatment for solid neoplasms.
Kanai
F
F
Second Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Shiratori
Y
Y
Yoshida
Y
Y
Wakimoto
H
H
Hamada
H
H
Kanegae
Y
Y
Saito
I
I
Nakabayashi
H
H
Tamaoki
T
T
Tanaka
T
T
Lan
K H
KH
Kato
N
N
Shiina
S
S
Omata
M
M
D000256:Adenoviridae / Q000235:genetics; D000595:Amino Acid Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000628:therapy*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D015774:Ganciclovir / Q000494:pharmacology; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005814:Genes, Viral; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D007763:Lac Operon; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000628:therapy*; D008969:Molecular Sequence Data; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000096:biosynthesis* / Q000235:genetics
D000509:alpha-Fetoproteins; D015500:Chloramphenicol O-Acetyltransferase; D013937:Thymidine Kinase; D015774:Ganciclovir