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Tunicamycin-induced Endoplasmic Reticulum Stress Upregulates the Expression of Pentraxin 3 in Human Retinal Pigment Epithelial Cells

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Purpose: To investigate the production of long pentraxin 3 (PTX3) in response to tunicamycin-induced endoplasmicreticulum (ER) stress and its role in ER stress-associated cell death, PTX3 expression was evaluatedin 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 treatmentby enzyme-linked immunosorbent assay. PTX3 protein and mRNA levels were estimated using western blotanalysis and real-time reverse transcription-polymerase chain reaction, respectively. Protein and mRNA levelsof CCAAT-enhancer-binding protein homologous protein (CHOP) and ARPE-19 cell viability were measuredin the presence of tunicamycin-induced ER stress in control or PTX3 small hairpin RNA (shRNA)-transfectedARPE-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-kB inhibitorabolished tunicamycin-induced PTX3 production. Decreased cell viability and prolonged protein and mRNAexpression 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-inducedER stress. Therefore, PTX3 could be an important protector of ER stress-induced cell death in human retinalpigment epithelial cells. Inositol-requiring enzyme 1α and the NF-kB signaling pathway may serve as potentialtargets for regulation of PTX3 expression in the retina. Therefore, their role in PTX3 expression needs to befurther investigated.

Abstract

Purpose: To investigate the production of long pentraxin 3 (PTX3) in response to tunicamycin-induced endoplasmicreticulum (ER) stress and its role in ER stress-associated cell death, PTX3 expression was evaluatedin 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 treatmentby enzyme-linked immunosorbent assay. PTX3 protein and mRNA levels were estimated using western blotanalysis and real-time reverse transcription-polymerase chain reaction, respectively. Protein and mRNA levelsof CCAAT-enhancer-binding protein homologous protein (CHOP) and ARPE-19 cell viability were measuredin the presence of tunicamycin-induced ER stress in control or PTX3 small hairpin RNA (shRNA)-transfectedARPE-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-kB inhibitorabolished tunicamycin-induced PTX3 production. Decreased cell viability and prolonged protein and mRNAexpression 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-inducedER stress. Therefore, PTX3 could be an important protector of ER stress-induced cell death in human retinalpigment epithelial cells. Inositol-requiring enzyme 1α and the NF-kB signaling pathway may serve as potentialtargets for regulation of PTX3 expression in the retina. Therefore, their role in PTX3 expression needs to befurther investigated.

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