Specificity of miR-378a-5p targeting rodent fibronectin
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- 作者：Fengqiong Liu ; Qing Lv ; William W. Du ; Haoran Li ; Xiangling Yang ; Danyang Liu ; Zhaoqun Deng ; Wenhua Ling ; Yaou Zhang ; Burton B. Yang
- 关键词：DMEM ; Dulbecco's modified Eagle's medium ; FBS ; fetal bovine serum ; GFP ; green fluorescent protein ; siRNA ; small interfering RNA ; miRNA ; microRNA ; 3&prime ; -UTR ; 3&prime ; -untranslated region
- 刊名：Biochimica et Biophysica Acta (BBA)/Molecular Cell Research
- 出版年：December, 2013
- 年：2013
- 卷：1833
- 期：12
- 页码：3272-3285
- 全文大小：4234 K
文摘
One criterion for microRNA identification is based on their conservation across species, and prediction of miRNA targets by empirical approaches using computational analysis relies on the presence of conservative mRNA 3鈥睻TR. Because most miRNA target sites identified are highly conserved across different species, it is not clear whether miRNA targeting is species-specific. To predict miRNA targeting, we aligned all available fibronectin 3鈥睻TRs and observed significant conservation of all 20 species. Twelve miRNAs were predicted to target most fibronectin 3鈥睻TRs, but rodent fibronectin showed potential binding sites specific for five different miRNAs. One of them, the miR-378a-5p, contained a complete matching seed-region for all rodent fibronectin, which could not be found in any other species. We designed experiments to test whether the species-specific targeting possessed biological function and found that expression of miR-378a-5p decreased cancer cell proliferation, migration, and invasion, resulting in inhibition of tumor growth. Silencing fibronectin expression produced similar effects as miR-378a-5p, while transfection with a construct targeting miR-378-5p produced opposite results. Tumor formation assay showed that enhanced expression of fibronectin in the stromal tissues as a background environment suppressed tumor growth, while increased fibronectin expression inside the tumor cells promoted tumor growth. This was likely due to the different signaling direction, either inside-out or outside-in signal. Our results demonstrated that species-specific targeting by miRNA could also exert functional effects. Thus, one layer of regulation has been added to the complex network of miRNA signaling.