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1 Free Radical and Radiation Biology Program, Department of Radiation Oncology, The University of Iowa, Iowa City, IA, USA
* To whom correspondence should be addressed. E-mail: larry-oberley{at}uiowa.edu.
Redox factor-1 (Ref-1/APE), a multifunctional DNA base excision repair and redox regulation enzyme, plays an important role in oxidative signaling, transcription factor regulation, and cell cycle control. We hypothesized that Ref-1 plays a regulatory role in smooth muscle cell (SMC) proliferation induced by platelet-derived growth factor (PDGF). ref-1 antisense oligodeoxynucleotides (AODN), which diminished the level of Ref-1 protein in SMC by approximately 50%, inhibited PDGF-BB-induced [3H]thymidine incorporation as compared with control oligodeoxynucleotides. ref-1 AODN inhibited PDGF-BB-induced S-entry by about 63%, which was overcome by overexpression of Ref-1 by adenoviral-mediated gene transfer. Overexpression of Ref-1 alone without PDGF enhanced SMC entry into S-phase. Furthermore, decreasing Ref-1 protein by treatment of SMC with Ref-1 AODN, or by immunodepletion of Ref-1 from nuclear extracts, inhibited PDGF-BB-induced AP-1 DNA binding activity. Chemical reduction restored the AP-1 DNA binding in Ref-1-depleted nuclear extracts. These results suggest that Ref-1 contributes to the regulation of PDGF-BB-stimulated cell cycle progression from G0/G1 to S in SMC, with one of the possible steps being redox-regulation of AP-1 by Ref-1 protein.
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