Lipopolysaccharide enhances DNA‑induced IFN‑β expression and autophagy by upregulating cGAS expression in A549 cells

Wang,Rong; Wang,Wei; Li,Aili; Wang,Yongqin; Jin,Junfei; Huang,Zhaoquan; Huang,Guojin

doi:10.3892/etm.2019.8001

Lipopolysaccharide enhances DNA‑induced IFN‑β expression and autophagy by upregulating cGAS expression in A549 cells

Authors:
- Rong Wang
- Wei Wang
- Aili Li
- Yongqin Wang
- Junfei Jin
- Zhaoquan Huang
- Guojin Huang
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Affiliations: Laboratory of Respiratory Disease, Affiliated Hospital of Guilin Medical University, Guilin, Guangxi 541001, P.R. China, China‑USA Lipids in Health and Disease Research Center, Guilin Medical University, Guilin, Guangxi 541001, P.R. China, Department of Pathology, Guilin Medical University, Guilin, Guangxi 541001, P.R. China
Published online on: September 13, 2019 https://doi.org/10.3892/etm.2019.8001
Pages: 4157-4164

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Abstract

Cyclic guanosine monophosphate‑adenosine monophosphate synthase (cGAS) is a newly identified cytosolic DNA sensor, but its function in lung epithelial cells is relatively unknown. In the present study, the effects of lipopolysaccharide (LPS) on the expression and function of cGAS in the A549 lung epithelial cell line was investigated. The cells were treated with LPS at different concentrations (e.g., 100, 200, 400 and 800 ng/ml), and the cGAS expression levels were examined via western blot analysis and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). The cells were pretreated with LPS, followed by E. coli DNA transfection using Lipofectamine® 3000. After 24 h, interferon (IFN)‑β production was measured using ELISA and the expression of the autophagic markers, microtubule‑associated proteins 1A/1B light chain 3 and sequestosome‑1, were determined using western blot analysis. The cells were also pretreated with either a toll‑like receptor (TLR) 4 inhibitor, a serine/threonine‑protein kinase TBK1 (TBK1) inhibitor or an nuclear factor (NF)‑κB inhibitor, followed by LPS treatment, and the cGAS expression levels were examined via western blot analysis and RT‑qPCR. The result showed that LPS treatment upregulated cGAS expression in a dose‑dependent manner. E. coli DNA treatment could induce IFN‑β production and autophagy via cGAS, which was enhanced by LPS pretreatment. The effect of LPS on cGAS expression was suppressed by treatment with a TLR4 inhibitor, a TBK1 inhibitor and an NF‑κB inhibitor. In conclusion, LPS enhances DNA‑induced IFN‑β production and autophagy by upregulating cGAS expression through the myeloid differentiation primary response protein MyD88‑independent TLR4 signaling pathway in A549 cells.

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