Fan Ruifeng, Yao Haidong, Zhao Xia, Cao Changyu, Yang Tianshu, Luan Yilin, Zhang Ziwei, Xu Shiwen
Department of Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People's Republic of China.
Biometals. 2016 Aug;29(4):679-89. doi: 10.1007/s10534-016-9943-3. Epub 2016 Jun 25.
The aim of the present study was to clarify the effect of Selenoprotein K (Selk) silencing on the mRNA expression of 25 selenoproteins in chicken myoblasts. The specific small interfering RNA (siRNA) for Selk gene was designed and transfected into chicken myoblasts. Post-transfection mRNA expression of 25 selenoproteins was determined at various time periods i.e., 24, 48 and 72 h. Moreover, based on the results of expression of 25 selenoproteins, correlation analysis and principal component analysis (PCA) were used for further analysis. The results showed that the designed siRNA effectively inhibited Selk expression (decreased by 20, 29 and 43 % on 24, 48 and 72 h, respectively) and the mRNA expression levels of the 23 selenoproteins were influenced by silencing Selk differently (P < 0.05). Time-dependent pattern of mRNA expression after siRNA treatment in three groups were found similar: one group including Gpx1, Gpx2, Gpx3, Gpx4, Txnrd1, Txnrd2, Txnrd3, Sepw1, Selh, Sepp1, Selo and Sepx1, another group including Sepn1, Sels, Selt, Selm and Sep15 and other group including Dio2 and Dio3. The results of correlation analysis showed that Gpx1, Gpx2, Gpx3, Gpx4, Dio1, Dio3, Sepn1, Sels, Sepw1, Selt, Selh, Sep15, Seli and Selu had a positive correlation with Selk, while Dio2 and Sepp1 had a negative correlation with Selk. PCA data also indicated that Txnrd1, Txnrd2, Dio2, Selpb, Sepp1and Selo may play special roles in response to Selk silencing. In summary, these results indicated that different selenoproteins possess and exhibits distinct responses to silencing of Selk in chicken myoblasts.
本研究旨在阐明硒蛋白K（Selk）沉默对鸡成肌细胞中25种硒蛋白mRNA表达的影响。设计了针对Selk基因的特异性小干扰RNA（siRNA）并将其转染到鸡成肌细胞中。在转染后的不同时间段，即24、48和72小时，测定25种硒蛋白的mRNA表达。此外，基于25种硒蛋白的表达结果，进行相关分析和主成分分析（PCA）以作进一步分析。结果表明，设计的siRNA有效抑制了Selk表达（在24、48和72小时分别降低了20%、29%和43%），并且23种硒蛋白的mRNA表达水平受到Selk沉默的不同影响（P<0.05）。发现三组中siRNA处理后mRNA表达的时间依赖性模式相似：一组包括Gpx1、Gpx2、Gpx3、Gpx4、Txnrd1、Txnrd2、Txnrd3、Sepw1、Selh、Sepp1、Selo和Sepx1，另一组包括Sepn1、Sels、Selt、Selm和Sep15，还有一组包括Dio2和Dio3。相关分析结果表明，Gpx1、Gpx2、Gpx3、Gpx4、Dio1、Dio3、Sepn1、Sels、Sepw1、Selt、Selh、Sep15、Seli和Selu与Selk呈正相关，而Dio2和Sepp1与Selk呈负相关。PCA数据还表明，Txnrd1、Txnrd2、Dio2、Selpb、Sepp1和Selo可能在对Selk沉默的反应中发挥特殊作用。总之，这些结果表明不同的硒蛋白对鸡成肌细胞中Selk沉默具有并表现出不同的反应。
