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Long non-coding (lnc)RNA profiling and the role of a key regulator lnc-PNRC2-1 in the transforming growth factor-1-induced epithelial-mesenchymal transition of CNE1 nasopharyngeal carcinoma cells

Journal of International Medical Research(2021)SCI 4区

Kunming Med Univ

Cited 0|Views21
Abstract
Objectives 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. Methods CNE1 cells were divided into transforming growth factor (TGF)-beta 1-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. Results TGF-beta 1-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 MYOM3 and reduced expression of EMT markers, resembling treatment with the TGF-beta 1 receptor inhibitor LY2109761. Conclusion Various lncRNAs participated indirectly in the TGF-beta 1-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.
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Nasopharyngeal carcinoma,epithelial&#8211,mesenchymal transition,long non-coding RNA,transforming growth factor-&#946,1,lnc-PNRC2-1,MYOM3
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要点】:研究确定了在鼻咽癌细胞CNE1上皮-间质转化(EMT)过程中起关键作用的lncRNA lnc-PNRC2-1,并探讨了其调控机制。

方法】:通过实时定量PCR和Western印迹分析TGF-β1诱导的EMT标志物的mRNA和蛋白质表达,利用RNA测序分析两组间的差异表达基因(DEGs),并进行基因本体和京都基因与基因组百科全书分析。

实验】:将CNE1细胞分为TGF-β1诱导的EMT组和对照组,通过RNA测序分析差异表达基因,并在lnc-PNRC2-1敲低后通过Western印迹重新评估EMT标志物表达,使用的数据集名称未在文中明确提及。实验结果表明,TGF-β1诱导的EMT表现为E-钙黏蛋白减少和波形蛋白、N-钙黏蛋白、Twist增加,lnc-PNRC2-1的敲低影响了EMT标志物的表达。