Wei Mengzhen, Tang Wenkui, Lv Danwei, Liu Mingxing, Wang Guodong, Liu Quanhui, Qin Liangshan, Huang Ben, Zhang Dandan
Guangxi Key Laboratory of Eye Health, Guangxi Academy of Medical Sciences, People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China.
Front Genet. 2023 Sep 5;14:1189487. doi: 10.3389/fgene.2023.1189487. eCollection 2023.
Long noncoding RNAs (lncRNAs) were important regulators affecting the cellular reprogramming process. Previous studies from our group have demonstrated that small molecule compounds can induce goat ear fibroblasts to reprogram into mammary epithelial cells with lactation function. In this study, we used lncRNA-Sequencing (lncRNA-seq) to analyze the lncRNA expression profile of cells before and after reprogramming (CK vs. 5i8 d). The results showed that a total of 3,970 candidate differential lncRNAs were detected, 1,170 annotated and 2,800 new lncRNAs. Compared to 0 d cells, 738 lncRNAs were significantly upregulated and 550 were significantly downregulated in 8 d cells. Heat maps of lncrnas and target genes with significant differences showed that the fate of cell lineages changed. Functional enrichment analysis revealed that these differently expressed (DE) lncRNAs target genes were mainly involved in signaling pathways related to reprogramming and mammary gland development, such as the Wnt signaling pathway, PI3K-Akt signaling pathway, arginine and proline metabolism, ECM-receptor interaction, and MAPK signaling pathway. The accuracy of sequencing was verified by real-time fluorescence quantification (RT-qPCR) of lncRNAs and key candidate genes, and it was also demonstrated that the phenotype and genes of the cells were changed. Therefore, this study offers a foundation for explaining the molecular mechanisms of lncRNAs in chemically induced mammary epithelial cells.
长链非编码RNA（lncRNAs）是影响细胞重编程过程的重要调节因子。我们团队之前的研究表明，小分子化合物可以诱导山羊耳成纤维细胞重编程为具有泌乳功能的乳腺上皮细胞。在本研究中，我们使用lncRNA测序（lncRNA-seq）分析重编程前后细胞的lncRNA表达谱（CK组与5i8 d组）。结果显示，共检测到3970个候选差异lncRNAs，其中1170个为注释lncRNAs，2800个为新的lncRNAs。与0 d细胞相比，8 d细胞中有738个lncRNAs显著上调，550个lncRNAs显著下调。具有显著差异的lncRNAs和靶基因的热图显示细胞谱系命运发生了变化。功能富集分析表明，这些差异表达（DE）lncRNAs的靶基因主要参与与重编程和乳腺发育相关的信号通路，如Wnt信号通路、PI3K-Akt信号通路、精氨酸和脯氨酸代谢、细胞外基质-受体相互作用以及MAPK信号通路。通过对lncRNAs和关键候选基因进行实时荧光定量（RT-qPCR）验证了测序的准确性，同时也证明了细胞的表型和基因发生了变化。因此，本研究为解释lncRNAs在化学诱导乳腺上皮细胞中的分子机制提供了基础。
