RRC ID 68498
Author Tanaka S, Iwai M, Harada Y, Morikawa T, Muramatsu A, Mori T, Okanoue T, Kashima K, Maruyama-Tabata H, Hirai H, Satoh E, Imanishi J, Mazda O.
Title Targeted killing of carcinoembryonic antigen (CEA)-producing cholangiocarcinoma cells by polyamidoamine dendrimer-mediated transfer of an Epstein-Barr virus (EBV)-based plasmid vector carrying the CEA promoter.
Journal Cancer Gene Ther
Abstract The present study reports a novel nonviral method to efficiently and specifically target carcinoembryonic antigen (CEA)-producing cholangiocarcinoma (CC) cells in vitro. Epstein-Barr virus (EBV)-based and conventional plasmid vectors were constructed that possess the beta-galactosidase (beta-gal) or herpes simplex virus-1 (HSV-1) thymidine kinase (Tk) genes as well as tandem repeats of the human genomic sequence -82 to -42 bp from the transcriptional start site of the CEA gene. The plasmids were transfected by means of polyamidoamine dendrimer into CEA-positive (HuCC-T1) or -negative cell lines. Transfection of the conventional plasmid vector with the CEA promoter and beta-gal gene resulted in a very low or undetectable level of marker gene expression even in the CEA-positive cell line. Transferring the HSV-1 Tk gene by conventional plasmid did not affect the susceptibility of HuCC-T1 cells to ganciclovir. In marked contrast, strong beta-gal expression was specifically obtained in HuCC-T1 cells by transfecting the EBV-based plasmid in which the CEA promoter and a ubiquitous promoter (SRalpha) are employed to drive the EBV-encoded nuclear antigen 1 (EBNA1) and beta-gal genes, respectively (pTES.beta). Furthermore, CEA-positive but not -negative tumor cells were rendered highly susceptible to ganciclovir when transfected with the EBV-based vector that carries the CEA promoter-EBNA1 and SRalpha-HSV-1 Tk genes (pTES.Tk). These results strongly suggest that the EBV-based plasmid vector/cationic polymer system (EBV/polyplex) equipped with the CEA promoter provides an efficient nonviral method for the targeted gene therapy of CEA-producing malignancies.
Volume 7(9)
Pages 1241-50
Published 2000-9-1
DOI 10.1038/sj.cgt.7700219
PMID 11023196
MeSH Carcinoembryonic Antigen / genetics Carcinoembryonic Antigen / metabolism* Cell Survival / drug effects Cholangiocarcinoma / metabolism Cholangiocarcinoma / pathology Cholangiocarcinoma / therapy* Colonic Neoplasms / metabolism Colonic Neoplasms / pathology Colonic Neoplasms / therapy* Coloring Agents DNA Primers / chemistry Epstein-Barr Virus Nuclear Antigens / genetics* Ganciclovir / pharmacology Genetic Therapy / methods Genetic Vectors* Humans Oxazines* Polyamines / therapeutic use* Polymerase Chain Reaction Promoter Regions, Genetic / genetics* Transfection / methods* Tumor Cells, Cultured / cytology Tumor Cells, Cultured / drug effects Xanthenes* beta-Galactosidase / metabolism
IF 4.534
Human and Animal Cells SSP-25(RCB1293)