Identification and characterization of NF-kappaB binding sites in human miR-1908 promoter

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• 作者：Qianhuining Kuang^a ; ^b ; ¹ ; ^{kuangqianhuining@126.com" class="auth_mail" title="E-mail the corresponding author} ; Jingyun Li^a ; ¹ ; Lianghui You^a ; Chunmei Shi^a ; Chenbo Ji^a ; Xirong Guo^a ; Meiyu Xu^b ; ; Yuhui Ni^a ;
• 关键词：miR-1908 ; NF-kappaB binding sites ; Promoter
• 刊名：Biomedicine & Pharmacotherapy
• 出版年：2015
• 出版时间：August 2015
• 年：2015
• 卷：74
• 期：Complete
• 页码：158-163
• 全文大小：1392 K

文摘

Previous studies reported that miR-1908 was highly expressed in mature human adipocytes. Adipokines tumor necrosis factor 伪 (TNF-伪) that was known to activate NF-kappaB signaling could affect the expression of miR-1908 in adipocytes. However, little is known about the underlying mechanisms.

Methods

In this study, we identified miR-1908 promoter using polymerase chain reaction (PCR) from human genomic DNA. Bioinformatic analysis was applied to predict the NF-kappaB binding sites in miR-1908 promoter. Real-time PCR, dual luciferase reporter assay, Mutagenesis analysis and electrophoretic mobility shift assay (EMSA) were performed to demonstrate the function of NF-kappaB binding sites in miR-1908 promoter.

Results

1243 bp miR-1908 promoter located in the intron of host gene fatty acid desaturase 1 (FADS1). Bioinformatic analysis revealed the presence of two putative NF-kappaB binding sites. TNF-伪 restricts miR-1908 expression in preadipocytes, and TNF-伪 decreases miR-1908 promoter activity in HEK293T cells. In addition, those two NF-kappaB transcription factor binding sites in miR-1908 promoter were functional.

Conclusion

Our findings demonstrated that miR-1908 has its own transcription unit, and revealed the transcriptional mechanisms of miR-1908 expression based on NF-kappaB signaling. This study offers a theoretical basis for understanding the transcriptional mechanism of miR-1908 expression and may provide a new strategy for obesity clinical therapy.