title: Utilization of variant-type of human alpha-fetoprotein promoter in gene therapy targeting for hepatocellular carcinoma.
abstract: We previously reported that the retroviral vector (LNAFW0.3TK) expressing the herpes simplex thymidine kinase (HSVtk) gene under the control of the 0.3 kb human alpha-fetoprotein (AFP) promoter provided the ganciclovir (GCV)-mediated cytotoxicity in the high AFP-producing (HuH-7) but not in the low AFP-producing (huH-1/cl.2) human hepatoma cells. In the present study, we constructed the retroviral vector (LNAFM0.3TK) in which the HSVtk gene expression is regulated by the variant-type of the 0.3 kb human AFP promoter with a G-to-A substitution at nucleotide -119, a point mutation responsible for hereditary persistence of human AFP and the vector was applied to three human hepatoma cell lines HuH-7, huH-1/cl.2 and intermediate AFP-producing cells (PLC/PRF/5). By the reporter gene transfection assay, the activity of the variant-type of the promoter was much higher than that of the wild-type of the promoter in both HuH-7 and huH-1/cl.2 cells. Consistent with this, LNAFM0.3TK infection could sensitize huH-1/cl.2 cells, as well as HuH-7 and PLC/PRF/5 cells to GCV, but did not affect cell growth of nonhepatoma cells (HeLa). In addition, the bystander effect was achieved more efficiently by LNAFM0.3TK infection than LNAFW0.3TK infection in HuH-7 cells. These results suggest that the variant-type of the human AFP promoter ensures the therapeutic gene expression in gene therapy particularly for the low AFP-producing hepatoma cells.
authors.affiliation: First Department of Internal Medicine, Nagasaki University School of Medicine, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Ishikawa
authors.forename: K
authors.initials: K
authors.lastname: Nakata
authors.forename: F
authors.initials: F
authors.lastname: Mawatari
authors.forename: T
authors.initials: T
authors.lastname: Ueki
authors.forename: S
authors.initials: S
authors.lastname: Tsuruta
authors.forename: A
authors.initials: A
authors.lastname: Ido
authors.forename: K
authors.initials: K
authors.lastname: Nakao
authors.forename: Y
authors.initials: Y
authors.lastname: Kato
authors.forename: N
authors.initials: N
authors.lastname: Ishii
authors.forename: K
authors.initials: K
authors.lastname: Eguchi
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000628:therapy*; D018390:Gene Targeting / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006801:Humans; D008113:Liver Neoplasms / Q000628:therapy*; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics*; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000235:genetics*
chemical_list: D000509:alpha-Fetoproteins; D013937:Thymidine Kinase