title: Hypoxia/hepatoma dual specific suicide gene expression plasmid delivery using bio-reducible polymer for hepatocellular carcinoma therapy.
abstract: Gene therapy is suggested as a promising alternative strategy of hepatocellular carcinoma (HCC, also called hepatoma) therapy. To achieve a successful and safe gene therapy, tight regulation of gene expression is required to minimize side-effects in normal tissues. In this study, we developed a novel hypoxia and hepatoma dual specific gene expression vector. The constructed vectors were transfected into various cell lines using bio-reducible polymer, PAM-ABP. First, pAFPS-Luc or pAFPL-Luc vector was constructed with the alpha-fectoprotein (AFP) promoter and enhancer for hepatoma tissue specific gene expression. Then, pEpo-AFPL-Luc was constructed by insertion of the erythropoietin (Epo) enhancer for hypoxic cancer specific gene expression. In vitro transfection assay showed that pEpo-AFPL-Luc transfected hepatoma cell increased gene expression under hypoxic condition. To confirm the therapeutic effect of dual specific vector, herpes simplex virus thymidine kinase (HSV-TK) gene was introduced for cancer cell killing. The pEpo-AFPL-TK was transfected into hepatoma cell lines in the presence of ganciclovir (GCV) pro-drug. Caspase-3/7, MTT and TUNEL assays elucidated that pEpo-AFPL-TK transfected cells showed significant increasing of death rate in hypoxic hepatoma cells compared to controls. Therefore, the hypoxia/hepatoma dual specific gene expression vector with the Epo enhancer and AFP promoter may be useful for hepatoma specific gene therapy.
authors.affiliation: Center for Controlled Chemical Delivery CCCD, Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, 84112, USA.
authors.forename: Hyun Ah
authors.initials: HA
authors.lastname: Kim
authors.forename: Kihoon
authors.initials: K
authors.lastname: Nam
authors.forename: Minhyung
authors.initials: M
authors.lastname: Lee
authors.forename: Sung Wan
authors.initials: SW
authors.lastname: Kim
mesh_terms: D000180:Acrylic Resins / Q000008:administration & dosage* / Q000737:chemistry; D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D050091:Dendrimers / Q000008:administration & dosage* / Q000737:chemistry; D004742:Enhancer Elements, Genetic; D004921:Erythropoietin / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D005814:Genes, Viral; D015316:Genetic Therapy; D057809:HEK293 Cells; D006801:Humans; D000860:Hypoxia / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D018139:Simplexvirus / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection / Q000379:methods; D000509:alpha-Fetoproteins / Q000235:genetics*
chemical_list: D000180:Acrylic Resins; D050091:Dendrimers; C554270:EPO protein, human; D000509:alpha-Fetoproteins; D004921:Erythropoietin; D004247:DNA; D008156:Luciferases; D013937:Thymidine Kinase
keywords: Bio-reducible polymer; Cancer hypoxia; Gene regulation; Hepatoma; Suicide gene therapy