Yang Jie, Feng Enzi, Ren Yanxin, Qiu Shun, Zhao Liufang, Li Xiaojiang
Head and Neck Tumor Research Center, The Third Affiliated Hospital of Kunming Medical University (Tumor Hospital of Yunnan Province & Yunnan Cancer Center), Kunming, Yunnan, China.
J Int Med Res. 2021 Mar;49(3):300060521996515. doi: 10.1177/0300060521996515.
To identify key long non-coding (lnc)RNAs responsible for the epithelial-mesenchymal transition (EMT) of CNE1 nasopharyngeal carcinoma cells and to investigate possible regulatory mechanisms in EMT.
CNE1 cells were divided into transforming growth factor (TGF)-β1-induced EMT and control groups. The mRNA and protein expression of EMT markers was determined by real-time quantitative PCR and western blotting. Differentially expressed genes (DEGs) between the two groups were identified by RNA sequencing analysis, and DEG functions were analyzed by gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses. EMT marker expression was re-evaluated by western blotting after knockdown of a selected lncRNA.
TGF-β1-induced EMT was characterized by decreased E-cadherin and increased vimentin, N-cadherin, and Twist expression at both mRNA and protein levels. Sixty lncRNA genes were clustered in a heatmap, and mRNA expression of 14 dysregulated lncRNAs was consistent with RNA sequencing. Knockdown of lnc-PNRC2-1 increased expression of its antisense gene and reduced expression of EMT markers, resembling treatment with the TGF-β1 receptor inhibitor LY2109761.
Various lncRNAs participated indirectly in the TGF-β1-induced EMT of CNE1 cells. Lnc-PNRC2-1 may be a key regulator of this and is a potential target to alleviate CNE1 cell EMT.
鉴定负责CNE1鼻咽癌细胞上皮-间质转化（EMT）的关键长链非编码（lnc）RNA，并研究EMT中可能的调控机制。
将CNE1细胞分为转化生长因子（TGF）-β1诱导的EMT组和对照组。通过实时定量PCR和蛋白质印迹法测定EMT标志物的mRNA和蛋白质表达。通过RNA测序分析鉴定两组之间的差异表达基因（DEG），并通过基因本体论和京都基因与基因组百科全书分析对DEG功能进行分析。在敲低选定的lncRNA后，通过蛋白质印迹法重新评估EMT标志物表达。
TGF-β1诱导的EMT的特征在于，在mRNA和蛋白质水平上，E-钙黏蛋白表达降低，波形蛋白、N-钙黏蛋白和Twist表达增加。60个lncRNA基因聚集在一个热图中，14个失调lncRNA的mRNA表达与RNA测序一致。敲低lnc-PNRC2-1可增加其反义基因的表达，并降低EMT标志物的表达，类似于用TGF-β1受体抑制剂LY2109761进行的治疗。
多种lncRNA间接参与TGF-β1诱导的CNE1细胞EMT。lnc-PNRC2-1可能是其中的关键调节因子，是减轻CNE1细胞EMT的潜在靶点。
