Effects of anticancer agents and scavengers on CMV-promoter-driven exogenous gene expression in genetically modified cells.
OBJECTIVE
This study aimed to investigate whether the levels of rsGFP mRNA and the fluorescence levels of cytomegalovirus (CMV)-promoter-driven rsGFP (red-shifted green fluorescent protein) could be changed by using anticancer agents and also to examine the effects of co-treatment with anticancer agents and scavengers.


METHODS
The pQBI25 vector, which encodes the CMV promoter and the cDNA for rsGFP, was transfected into FR cells (rat skin fibroblast cell line). FR-pQBI25 cells were then exposed to doxorubicin, 5-fluorouracil, methotrexate or paraquat with or without scavengers such as N-acetyl cysteine (NAC) and edaravone for 48 h.


RESULTS
The levels of rsGFP mRNA were found to be significantly higher following doxorubicin, 5-fluorouracil and paraquat treatment but were not changed by methotrexate. These levels of rsGFP mRNA were found to be significantly lower after paraquat/edaravone co-treatment compared with paraquat alone. The fluorescence levels of rsGFP were found to be significantly higher following doxorubicin and paraquat treatment but were not changed by 5-fluorouracil and methotrexate. The levels were also found to be significantly lower after paraquat/edaravone co-treatment compared with paraquat alone and also after doxorubicin/NAC co-treatment compared with doxorubicin alone.


CONCLUSIONS
These findings suggest that CMV-promoter-driven exogenous gene expression may be partly regulated by reactive oxygen species.
Kinoshita
Atsushi
A
Division of Drug Informatics, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Himeji, Hyogo, Japan.
Kobayashi
Daisuke
D
Saitoh
Yukiya
Y
Komada
Fusao
F
D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D002461:Cell Line, Transformed; D003587:Cytomegalovirus / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism*; D005453:Fluorescence; D016166:Free Radical Scavengers / Q000008:administration & dosage / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D014162:Transfection
D000970:Antineoplastic Agents; D016166:Free Radical Scavengers; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins