Experimental and Molecular Medicine

Korean Society for Biochemistry and Molecular Biongy (생화학분자생물학회)
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NF-$\kappa$B decoy potentiates the effects of radiation on vascular smooth muscle cells by enhancing apoptosis

Zhang, Shu-Ying;Park, Kyung-Woo;Oh, Se-Il;Cho, Hyun-Ju;Cho, Hyun-Jai;Park, Jin-Shik;Cho, Young-Seok;Koo, Bon-Kwon;Chae, In-Ho;Choi, Dong-Joo;Kim, Hyo-Soo;Lee, Myoung-Mook

  • Published : 2005.02.28
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Abstract

NF-kB promotes cell survival against external stress such as radiation. We examined whether NF-kB decoy transfection enhances the antiproliferative effects of radiation on vascular smooth muscle cells (VSMCs) in vitro. The irradiation induced activation or nuclear translocation of NF-kB p65 in VSMCs was confirmed by immunofluorescence. NF-kB decoy transfection resulted in inhibition of the radiation-induced NF-kB activation in VSMCs and the subsequent reduction of transcription and translocation of ICAM, iNOS, and TNF-a, downstream molecules under the control of NF-kB. By using MTT assay, NF-kB decoy augmented the antiproliferative effects of radiation, where the effect of low dose radiation (2 and 8-Gy) of the cells transfected with NF-kB decoy was equivalent to the high dose (16-Gy) irradiated non-transfected cells at 48 h after irradiation: 1.06±0.16, 1.11±0.22, 1.20±0.25, respectively. The decrease in proliferation and survival of the radiation treated cells by flow cytometry analysis showed that NF-kB inhibition did not show any additive effects on the cell cycle of the irradiated VSMCs, while apoptosis was significantly increased after NF-kB decoy transfection in the irradiated VSMCs (apoptosis fraction: 13.33±2.08% vs. 26.29±7.43%, for radiation only vs. radiation+NF-kB decoy transfection, P＜0.05). In addition, at 48 h, NF-kB decoy transfection dose dependently (10 mM vs. 20 mM) inhibited proliferation of 16Gy-irradiated VSMCs, and showed greater antiproliferative efficacy than 100 mM sulfasalazine, a specific NF- kB inhibitor. These results indicate that NF-kB inhibition reduces proliferation and survival of irradiated VSMCs, likely by increased apoptosis rather than additive cell cycle arrest and suggest the possibility of adjunctive gene therapy using NF-kB decoy to improve efficacy and to decrease the adverse effects of intracoronary radiation therapy.

Keywords

References

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