LncRNA DANCR调控非小细胞肺癌细胞的凋亡和上皮间质转化-现代医学

LncRNA DANCR调控非小细胞肺癌细胞的凋亡和上皮间质转化

单位：空军军医大学第二附属医院肿瘤科, 陕西西安 710038

关键词：非小细胞肺癌细胞 DANCR FOXO1/mTOR信号通路细胞凋亡上皮间质转化

分类号：R734.2

出版年·卷·期（页码）：2020·48·第一期（75-82）

摘要：

目的： 探讨促癌基因lncRNA DANCR在非小细胞肺癌（NSCLC）细胞的凋亡和上皮间质转化（EMT）中的分子机制。方法： 培养NSCLC细胞系A549；对细胞使用siRNA沉默DANCR，qRT-PCR 检测DANCR水平变化，Western blot检测DANCR靶基因FOXO1与其下游信号mTOR的表达；使用siRNA共沉默DANCR和FOXO1，Western blot检测FOXO1与其下游信号mTOR，VEGF，EMT诱导因子Twist1，EMT标记蛋白E-cadherin、vimentin 的表达，Caspase3活性检测和cleaved-caspase3水平变化评估A549细胞的凋亡水平，Transwell细胞侵袭实验和ELISA检测外分泌VEGF水平变化评估细胞侵袭能力。方法： 沉默DANCR显著性促进A549细胞中FOXO1和mTOR表达（P<0.05）；与单独沉默DANCR相比，共沉默DANCR和FOXO1显著性减缓沉默DANCR对Caspase3活性和cleaved-caspase3水平的促进作用；沉默DANCR对细胞侵袭能力胞外VEGF和胞内VEGF表达、EMT诱导因子Twist1表达和EMT标记物的表达的抑制作用均被共沉默DANCR和FOXO1显著性逆转（P<0.05）。方法： LncRNA DANCR通过FOXO1/mTOR信号通路调节NSCLC的凋亡和EMT。

Objective: To explore the molecular mechanism of DANCR regulating apoptosis and epithelial mesenchymal transformation (EMT) in non-small cell lung cancer (NSCLC) cells. Methods: The NSCLC cell line A549 was cultured. We silenced DANCR using siRNAs in the cells. qRT-PCR was used to detect changes in DANCR levels. Western blot was used to detect the expression of DANCR target gene FOXO1 and the downstream signal mTOR. Further, DANCR and FOXO1 were co-silenced using siRNAs. Western blot was used to detect the expression of FOXO1, mTOR and VEGF, EMT-inducible factor Twist1, and E-cadherin and vimentin, the EMT markers. Apoptosis of A549 cells were assessed by caspase3 activity and cleaved-caspase3 level changes. Furthermore, transwell invasion assay and ELISA assay were used to assess cell invasion ability the exocrine VEGF levels, respectively. Results: DANCR knockdown significantly promoted the expression of FOXO1 and mTOR in A549 cells (P<0.05). Co-silencing of DANCR and FOXO1 significantly slowed the promotion of Caspase3 activity and cleaved-caspase3 levels by silencing DANCR compared to DANCR alone. The inhibitory effects of silencing DANCR on cell invasion, extracellular VEGF levels, and intracellular VEGF, Twist1 and EMT markers were significantly reversed by co-silencing of DANCR and FOXO1. Conclusion: DANCR regulates apoptosis and EMT via FOXO1/mTOR signaling pathway in NSCLC cells.

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