Irradiation, cisplatin, and 5-azacytidine upregulate cytomegalovirus promoter in tumors and muscles: implementation of non-invasive fluorescence imaging.
Mol Imaging Biol
; 13(1): 43-52, 2011 Feb.
Article
in En
| MEDLINE
| ID: mdl-20396957
ABSTRACT
PURPOSE:
The cytomegalovirus (CMV) promoter is one of the most commonly used promoters for expression of transgenes in mammalian cells. The aim of our study was to evaluate the role of methylation and upregulation of the CMV promoter by irradiation and the chemotherapeutic agent cisplatin in vivo using non-invasive fluorescence in vivo imaging. PROCEDURES Murine fibrosarcoma LPB and mammary carcinoma TS/A cells were stably transfected with plasmids encoding CMV and p21 promoter-driven green fluorescent protein (GFP) gene. Solid TS/A tumors were induced by subcutaneous injection of fluorescent tumor cells, while leg muscles were transiently transfected with plasmid encoding GFP under the control of the CMV promoter. Cells, tumors, and legs were treated either by DNA methylation inhibitor 5-azacytidine, irradiation, or cisplatin. GFP expression was determined using a fluorescence microplate reader in vitro and by non-invasive fluorescence imaging in vivo.RESULTS:
Treatment of cells, tumors, and legs with 5-azacytidine (re)activated the CMV promoter. Furthermore, treatment with irradiation or cisplatin resulted in significant upregulation of GFP expression both in vitro and in vivo.CONCLUSIONS:
Observed alterations in the activity of the CMV promoter limit the usefulness of this widely used promoter as a constitutive promoter. On the other hand, inducibility of CMV promoters can be beneficially used in gene therapy when combined with standard cancer treatment, such as radiotherapy and chemotherapy.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Azacitidine
/
Up-Regulation
/
Promoter Regions, Genetic
/
Cisplatin
/
Genes, Immediate-Early
/
Cytomegalovirus
/
Muscles
Limits:
Animals
Language:
En
Journal:
Mol Imaging Biol
Journal subject:
BIOLOGIA MOLECULAR
/
DIAGNOSTICO POR IMAGEM
Year:
2011
Document type:
Article
Affiliation country: