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Nuclear Factor-B Is Required for Tumor Necrosis Factor--Induced Manganese Superoxide Dismutase Expression in Human Endometrial Stromal Cells

Department of Obstetrics and Gynecology, Yamaguchi University School of Medicine, Minamikogushi 1-1-1, Ube 755-8505, Japan

Address all correspondence and requests for reprints to: Norihiro Sugino, M.D., Department of Obstetrics and Gynecology, Yamaguchi University School of Medicine, Minamikogushi 1-1-1, Ube 755-8505, Japan. E-mail:; . obgyn{at}po.cc.yamaguchi-u.ac.jp

Abstract

We recently found that manganese superoxide dismutase (Mn-SOD) is up-regulated by TNF at the transcription level in human endometrial stromal cells (ESC) and that TNF- induced Mn-SOD expression is mediated by protein kinase C (PKC)-dependent phosphorylation. This study was undertaken to investigate whether nuclear factor-B (NF-B), a transcription factor, is involved in Mn-SOD induction by TNF or PKC in human ESC. Electrophoretic mobility shift assay revealed that TNF (1 ng/ml) and phorbol 12-myristate 13-acetate (TPA; 0.4 µM), PKC activator, caused marked increases in nuclear NF-B DNA binding activity. Secondly, ESC were incubated with MG132 (proteasome inhibitor) or SN50 (inhibitor of translocation of NF-B into the nucleus) in the presence of TNF or TPA. TNF and TPA significantly increased Mn-SOD activities and Mn-SOD mRNA levels, and those effects were completely inhibited by MG132 and SN50. TNF alone caused no effect on cell viability, but in the presence of MG132, TNF significantly decreased cell viability. This inhibitory effect of MG132 was blocked by simultaneous addition of N-acetyl-L-cysteine, an antioxidant. In conclusion, the present study showed the involvement of NF-B in Mn-SOD induction by TNF or PKC in human ESC. This phenomenon could be a self-defense system of ESC against TNF-mediated oxidative stress.

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