Herp depletion inhibits zearalenone-induced cell death in RAW 264.7 macrophages. (April 2016)

• Record Type: Journal Article
• Authors: Chen, Fenglei; Lin, Pengfei; Wang, Nan; Yang, Diqi; Wen, Xin; Zhou, Dong; Wang, Aihua; Jin, Yaping
• Is Part Of: Toxicology in vitro. Volume 32(2016)
• Page Start: 115
• Page End: 122
• Publication Date: 2016-04
• Subjects: Atg autophagy-related proteins -- DMEM Dulbecco's modified Eagle's medium -- DMSO dimethyl sulfoxide -- ER endoplasmic reticulum -- ERAD ER-associated protein degradation -- FBS fetal bovine serum -- Fisher LSD Fisher's least significant different test -- GS glucose starvation -- Herp homocysteine-responsive ER-resident protein -- LC3 microtubule-associated protein 1 light chain 3 -- LPS lipopolysaccharide -- MTT 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide assay -- poly-Ub poly-ubiquitin -- PVDF polyvinylidene difluoride filter -- RIPA radioimmunoprecipitation -- shHerp Herp-specific shRNA constructs -- shNC a negative control shRNA -- shRNAs short hairpin interfering RNAs -- Tg thapsigargin -- ULD N-terminal ubiquitin-like domain -- UPR unfolded protein response -- VSVG vesicular stomatitis virus -- ZEA zearalenone; Herp -- Autophagy -- Zearalenone -- Cell death -- LC3; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2015.12.014
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Herp is an endoplasmic reticulum (ER) membrane protein and strongly induced by the ER stress that not only participates in the unfolded protein response (UPR) under the ER stress, but also in cell autophagy under glucose starvation (GS). However, we do not know whether Herp plays any roles in other responses, such as zearalenone (ZEA). In this study, we constructed recombinant lentiviral vectors for Herp shRNA expression and generated stable Herp knockdown RAW 264.7 macrophages. Flow cytometry analysis showed Herp depletion could inhibit cell death induced by ZEA. Western blot analysis revealed that Herp depletion could up-regulate autophagy-related protein LC3-I conversion into LC3-II and the expression of ER stress-related protein CHOP. These results suggest that Herp depletion inhibits cell death by up-regulating autophagy. Highlights: ZEA up-regulated ER stress-related proteins Herp expression and autophagy-related protein LC3-I conversion to LC3-II in dose- and time-dependent manners. Herp depletion up-regulated LC3-I conversion to LC3-II treated with ZEA. Herp depletion had a protective role in ZEA-induced cell death via up-regulating autophage.