Tunicamycin-induced Endoplasmic Reticulum Stress Upregulates the Expression of Pentraxin 3 in Human Retinal Pigment Epithelial Cells.
10.3341/kjo.2016.30.6.468
- Author:
Narae HWANG
1
;
Min Young KWON
;
Jae Bong CHA
;
Su Wol CHUNG
;
Je Moon WOO
Author Information
1. School of Biological Sciences, College of Natural Sciences, University of Ulsan, Ulsan, Korea.
- Publication Type:Original Article
- Keywords:
Endoplasmic reticulum stress;
Human retinal pigment epithelial cells;
Inositol-requiring enzyme 1α;
Pentraxin 3
- MeSH:
Anti-Bacterial Agents/pharmacology;
Apoptosis;
Blotting, Western;
C-Reactive Protein/biosynthesis/*genetics;
Cells, Cultured;
Endoplasmic Reticulum Stress/*drug effects/genetics;
Enzyme-Linked Immunosorbent Assay;
*Gene Expression Regulation;
Humans;
Polymerase Chain Reaction;
RNA, Messenger/*genetics;
Retinal Pigment Epithelium/*metabolism/pathology;
Serum Amyloid P-Component/biosynthesis/*genetics;
Tunicamycin/*pharmacology
- From:Korean Journal of Ophthalmology
2016;30(6):468-478
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: To investigate the production of long pentraxin 3 (PTX3) in response to tunicamycin-induced endoplasmic reticulum (ER) stress and its role in ER stress-associated cell death, PTX3 expression was evaluated in the human retinal pigment epithelial cell line, ARPE-19. METHODS: PTX3 production in ARPE-19 cells was analyzed in the absence or presence of tunicamycin treatment by enzyme-linked immunosorbent assay. PTX3 protein and mRNA levels were estimated using western blot analysis and real-time reverse transcription-polymerase chain reaction, respectively. Protein and mRNA levels of CCAAT-enhancer-binding protein homologous protein (CHOP) and ARPE-19 cell viability were measured in the presence of tunicamycin-induced ER stress in control or PTX3 small hairpin RNA (shRNA)-transfected ARPE-19 cells. RESULTS: The protein and mRNA levels of PTX3 were found to be significantly increased by tunicamycin treatment. PTX3 production was significantly decreased in inositol-requiring enzyme 1α shRNA-transfected ARPE-19 cells compared to control shRNA-transfected cells. Furthermore, pretreatment with the NF-κB inhibitor abolished tunicamycin-induced PTX3 production. Decreased cell viability and prolonged protein and mRNA expression of CHOP were observed under tunicamycin-induced ER stress in PTX3 shRNA transfected ARPE-19 cells. CONCLUSIONS: These results suggest that PTX3 production increased in the presence of tunicamycin-induced ER stress. Therefore, PTX3 could be an important protector of ER stress-induced cell death in human retinal pigment epithelial cells. Inositol-requiring enzyme 1α and the NF-κB signaling pathway may serve as potential targets for regulation of PTX3 expression in the retina. Therefore, their role in PTX3 expression needs to be further investigated.
