GRP78 silencing enhances hyperoxia-induced alveolar epithelial cell apoptosis via CHOP pathway

Lu,Hong‑Yan; Chen,Xiao‑Qing; Tang,Wei; Wang,Qiu‑Xia; Zhang,Jie

doi:10.3892/mmr.2017.6681

GRP78 silencing enhances hyperoxia-induced alveolar epithelial cell apoptosis via CHOP pathway

Authors:
- Hong‑Yan Lu
- Xiao‑Qing Chen
- Wei Tang
- Qiu‑Xia Wang
- Jie Zhang
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Affiliations: Department of Pediatrics, The Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212001, P.R. China, Department of Pediatrics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China
Published online on: June 2, 2017 https://doi.org/10.3892/mmr.2017.6681
Pages: 1493-1501

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Abstract

Hyperoxia is one of the primary causes of bronchopulmonary dysplasia, which may occur in premature infants following supplemental oxygen therapy. Glucose regulated protein 78 (GRP78), which is a molecular chaperone located in the lumen of the endoplasmic reticulum (ER), has been reported to regulate hyperoxia‑associated ER stress. The role of GRP78 in lung epithelial cells during hyperoxia remains to be elucidated. In the present study, the A549 cultured human lung epithelial cell line was exposed to hyperoxic conditions, and then transfected with short interfering (si)RNA targeted to GRP78. siRNA or pEGFP‑N1 plasmid were used to knockdown or overexpress specific genes, reverse transcription-quantitative polymerase chain reaction and western blot analysis were used to detect RNA and protein levels of gene expression, and flow cytometry was used to detect apoptosis. The expression levels of ER stress‑associated genes were determined, and a significant increase in C/EBP homologous protein (CHOP) expression and apoptosis of A549 cells was observed, following GRP78 knockdown. The overexpression of CHOP downregulated B‑cell lymphoma (Bcl)‑2 expression levels, upregulated BCL2 associated X (Bax), and increased apoptosis of A549 cells under conditions of hyperoxia. CHOP knockdown demonstrated the opposite effect on Bcl‑2 and Bax expression levels. These results suggested that GRP78 silencing promoted lung epithelial cell apoptosis during hyperoxia, via regulation of the CHOP pathway.

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