Regulation of exogenous gene expression by superoxide

• Published in: Pharmaceutical research Vol. 23; no. 11; pp. 2536 - 2541
• Main Authors: KINOSHITA, Atsushi, KOBAYASHI, Daisuke, SAITOH, Yukiya, TANABE, Naoto, UCHINO, Katsuyoshi, NISHIGUCHI, Kohshi, OKUMURA, Katsuhiko, KOMADA, Fusao
• Format: Journal Article
• Language: English
• Published: New York, NY Springer 01.11.2006; Springer Nature B.V
• Subjects: Animals; Biological and medical sciences; Cell Line; Cytomegalovirus; Cytomegalovirus - genetics; Gene expression; Gene Expression Regulation; Gene therapy; General pharmacology; Genes; Genes, fos; Genes, jun; Genetic Therapy; Medical sciences; Oxidation; Paraquat - pharmacology; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Rats; Regulation; Ribonucleic acid; RNA; RNA, Messenger - analysis; Superoxide Dismutase - biosynthesis; Superoxides - pharmacology; Terminal Repeat Sequences; Cytomegalovirus; Exogenous; Pharmaceutical technology; Herpesviridae; Gene expression; Betaherpesvirinae; AP-1; Infection; Virus; Hyperoxides; Gene; long terminal repeat; gene regulation; Viral disease; Regulation; superoxide
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-006-9076-4

Regulation of gene expression after gene introduction is a problematic aspect of gene therapy. Transcription regulates gene-specific transcriptional factors, which bind to regulatory regions in the promoter. The cytomegalovirus long terminal repeat (CMV-LTR) has a TPA response element (TRE) as a binding site for activator protein 1 (AP-1), which is induced by oxidative stress. The purpose of this study was to regulate exogenous gene expression in a vector with CMV-LTR using oxygen radicals. We used two plasmids (1) pQBI25 encoding CMV-LTR and red-shift green fluorescent protein (rsGFP) cDNA and (2) pRc/CMV-SOD encoding CMV-LTR and human Cu, Zn-superoxide dismutase (SOD) cDNA. FR cells were transfected with pQBI25 (FR-pQBI25 cells), and L2 cells were transfected with pRc/CMV-SOD (L2-pRc/CMV-SOD cells). Each type of cell was exposed to oxygen radicals using paraquat for 24 h. Levels of c-fos, c-jun and rsGFP mRNAs were determined using reverse transcription polymerase chain reaction (RT-PCR). Levels of rsGFP protein were measured by fluorometry. Total SOD activity was measured using the nitrite method. Levels of c-fos, c-jun (AP-1 composition protein) and rsGFP mRNA were induced significantly by oxygen radical exposure in FR-pQBI25 cells. A positive correlation was observed between levels of c-fos mRNA and rsGFP mRNA and also between levels of c-jun mRNA and rsGFP mRNA. Levels of rsGFP protein were also induced significantly. Total SOD activity was induced significantly by oxygen radical exposure in L2-pRc/CMV-SOD cells. This study suggests that gene expression driven by the CMV- LTR promoter may be regulated by oxygen radicals.