RRC ID 4788
Author Nagayama Y, Nishihara E, Namba H, Yokoi H, Hasegawa M, Mizuguchi H, Hayakawa T, Hamada H, Yamashita S, Niwa M.
Title Targeting the replication of adenovirus to p53-defective thyroid carcinoma with a p53-regulated Cre/loxP system.
Journal Cancer Gene Ther.
Abstract In this article, we evaluated the feasibility of the restricted replication-competent adenoviruses for treatment of anaplastic thyroid carcinomas (ATCs), which are very aggressive and difficult to treat. Because ATCs very often harbor p53 mutations, we used wt-p53 as a regulatory factor to restrict virus replication and cytopathic effect to p53-mutated cells. The recently reported "gene inactivation strategy" using p53-regulated Cre/loxP system was employed; this system consists of two recombinant adenoviruses. One has an expression unit of the synthetic p53 - responsive promoter and the Cre recombinase gene (Axyp53RECre), and another contains two expression units; the first consists of E1A gene flanked by a pair of loxP sites downstream of the constitutive CAG promoter and the second E1B19K gene under the control of the CMV promoter (AdCALE1AL). We expected that coinfection of these two adenoviruses into the cells with wt-p53 would lead to expression of the Cre, which excises E1A gene and switches off E1A expression resulting in no virus replication, whereas in the cells with mutant p53 E1A could be expressed that leads to virus replication and cell lysis. Our in vitro data demonstrate that although infection of AdCALE1AL alone led to E1A expression, viral replication and cytolysis in all the thyroid cells examined irrespective of their p53 status, the double infection did so in FRO cells (p53-null ATC) but not in FRO cells stably expressing wt-p53 and normal thyroid cells with wt-p53. These data indicate that our double infection method may have a potential for treatment of ATC and probably also other p53-defective cancer cells.
Volume 8(1)
Pages 36-44
Published 2001-1
DOI 10.1038/sj.cgt.7700276
PMID 11219492
MeSH Adenovirus E1A Proteins / metabolism Adenoviruses, Human / physiology* Antiviral Agents / adverse effects Cell Division / drug effects Ganciclovir / adverse effects Gene Expression Genetic Therapy / methods* Humans Immunoblotting Integrases / metabolism* Lac Operon / physiology Recombinant Proteins / metabolism Thyroid Neoplasms / metabolism Thyroid Neoplasms / pathology Thyroid Neoplasms / therapy* Thyroid Neoplasms / virology Tumor Cells, Cultured Tumor Suppressor Protein p53 / genetics* Viral Proteins* Virus Replication* / physiology beta-Galactosidase / metabolism
IF 4.044
Times Cited 24
WOS Category MEDICINE, RESEARCH & EXPERIMENTAL BIOTECHNOLOGY & APPLIED MICROBIOLOGY ONCOLOGY GENETICS & HEREDITY
Resource
DNA material pCALwL (RDB01679) AxCALNLNZ (RDB01750).