RNA-Seq reveals the role of miR-199a-3p in regulating inflammation of bovine mammary epithelial cells

Purpose: to detect expression profile of differentially expressed mRNAs during bovine mammary epithelial cells transfected with miR-199a-3p mimic or negative control (NC) mimic in vitro. Methods: bovine mammary epithelial cells were transfected with miR-199a-3p mimic or negative control (NC) mimic to assess the expression profiles of mRNAs using RNA-seq. Results: there were 140 DEGs (109 up-regulated and 31 down-regulated) in the miR-199a-3p overexpression group compared with the negative control group. Conclusion: the overexpression of miR-199a-3p resulted in differential expression of 140 genes in bMECs. The functional annotation results of these DEGs suggested that miR-199a-3p may be involved in the inflammatory and immune responses of bMECs by regulating PI3K-Akt signaling pathway, TGF-beta signaling pathway and IL-17 signaling pathway. Overall design: Bovine mammary epithelial cells were inoculated and cultured in 6-well cell culture plates. When cell confluence reached 60-70%, miR-199a-3p mimic and NC mimic (final concentration 50nM) were transiently transfected into cells. Subsequently, all samples were used to perform the RNA-seq.

研究目的：本数据集旨在体外检测转染miR-199a-3p模拟物（miR-199a-3p mimic）或阴性对照（NC）模拟物的牛乳腺上皮细胞（下文简称bMECs）中差异表达mRNA的表达谱。
实验方法：将bMECs转染miR-199a-3p模拟物或阴性对照模拟物，通过RNA测序（RNA-seq）分析mRNA的表达谱。
实验结果：与阴性对照组相比，miR-199a-3p过表达组中共存在140个差异表达基因（DEGs），其中上调基因109个，下调基因31个。
研究结论：miR-199a-3p过表达可导致bMECs中140个基因出现差异表达。对上述差异表达基因进行功能注释后发现，miR-199a-3p可能通过调控PI3K-Akt信号通路、转化生长因子β（TGF-β）信号通路以及白细胞介素17（IL-17）信号通路，参与bMECs的炎症与免疫应答。
整体实验设计：将bMECs接种于6孔细胞培养板中进行培养，待细胞汇合度达到60%~70%时，将终浓度为50nM的miR-199a-3p模拟物与阴性对照模拟物分别瞬时转染至细胞中。随后收集所有样本进行RNA测序（RNA-seq）。