Differentially expressed lncRNAs and mRNAs in primary human airway granulation fibroblasts before and after β-elemene treatment for 48h

There is an urgent need to identify effective medications to inhibit the abnormal airway granulation fibroblast activity, which plays a critical role in granulation tissue hyperplasia and scar formation during wound-healing after endobronchial intervention in the human airway, so as to prevent benign airway restenosis. Our previous study shown that β-elemene, a widely used effective ingredient of traditional Chinese medicine, effectively inhibited proliferation and survival of primary human airway granulation fibroblasts (PHAGF) in vitro, and promotes granulation tissue degeneration in vivo. To further investigate the mechanistic basis of these effects, we have employed LncRNA Microarray expression profiling as a discovery platform to identify differentially expressed lncRNAs and mRNAs in β-Elemene-treated PHAGF, thus enriching β-Elemene-altered pathway associated with cell proliferation and apoptosis, and thereby constructing a β-elemene-altered competitive endogenous RNAs (ceRNA) network that modulates this pathway. The results revealed ILK/Akt pathway, containing the upstream factor, ILK, the downstream cell cycle gene, Cyclin D1, and the anti-apoptosis gene, Bcl-2, were suppressed in β-Elemene-treated PHAGF. LncRNA-MIR143HG, which acts as a miR-1275 ceRNA to modulate ILK expression was also suppressed. Furhter validation via wet-lab experiments revealed that β-Elemene down-regulated LncRNA MIR143HG in PHAGF to reduce the competing sponging effect of MIR143HG on microRNA miR-1275, which targets upstream gene ILK in ILK/Akt pathway, thereby inducing the fibroblasts cell cycle arrest and apoptosis by suppressing ILK/Akt pathway. LncRNAs and mRNAs expressions in third-generation primary fibroblasts cultured from human airway hyperplastic granulation tissue resulting from prolonged intubation were measured with or without 160ug/ml β-elemene treatment for 48h, respectively. Three independent experiments were performed with or without β-elemene treatment.