Related Experiment Video
Updated: Jun 24, 2026
Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice
Published on: June 13, 2012
Effects of anticancer agents and scavengers on CMV-promoter-driven exogenous gene expression in genetically modified
Atsushi Kinoshita1, Daisuke Kobayashi, Yukiya Saitoh
1Division of Drug Informatics, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Himeji, Hyogo, Japan.
Objectives:
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.
Key Findings:
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.
Insights
Anticancer agents like doxorubicin and paraquat increased red-shifted green fluorescent protein (rsGFP) mRNA and fluorescence. Co-treatments with scavengers like N-acetyl cysteine (NAC) and edaravone modulated these effects, suggesting reactive oxygen species regulate gene expression.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Gene expression regulation is crucial in cellular processes.
- Cytomegalovirus (CMV) promoter is widely used for driving exogenous gene expression.
- Reactive oxygen species (ROS) can influence cellular functions and gene expression.
Purpose of the Study:
- To investigate the impact of anticancer agents on rsGFP mRNA and fluorescence levels driven by a CMV promoter.
- To examine the modulatory effects of ROS scavengers on these changes.
- To explore the potential role of ROS in regulating CMV-promoter-driven gene expression.
Main Methods:
- Transfection of FR cells with a pQBI25 vector encoding CMV promoter and rsGFP.
- Exposure of transfected cells to doxorubicin, 5-fluorouracil, methotrexate, or paraquat.
- Co-treatment experiments with ROS scavengers N-acetyl cysteine (NAC) and edaravone.
Main Results:
- Doxorubicin, 5-fluorouracil, and paraquat significantly increased rsGFP mRNA levels.
- Methotrexate did not alter rsGFP mRNA or fluorescence.
- Doxorubicin and paraquat significantly increased rsGFP fluorescence.
- Co-treatment with paraquat/edaravone or doxorubicin/NAC decreased rsGFP levels compared to agent alone.
Conclusions:
- CMV-promoter-driven exogenous gene expression is potentially regulated by ROS.
- Anticancer agents can modulate gene expression through ROS-dependent pathways.
- ROS scavengers can mitigate the effects of certain agents on gene expression.
