Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
Doctoral School of Molecular, Cell and Immune Biology, University of Debrecen, Debrecen, Hungary.
PLoS One. 2022 Jun 15;17(6):e0269936. doi: 10.1371/journal.pone.0269936. eCollection 2022.
Mutation-induced protein misfolding of pancreatic secretory enzymes and consequent endoplasmic reticulum stress can cause chronic pancreatitis. A recent study revealed that cigarette smoke also increases the risk of the disease through endoplasmic reticulum stress. Here, we investigated the cumulative cellular effect of the G233E misfolding human pancreatic lipase variant and hydroquinone; a main toxic constituent of cigarette smoke, using mammalian cell lines. We found that hydroquinone reduces cell viability on a dose-dependent manner through programmed cell death, and diminishes lipase secretion without affecting its expression. Interestingly, hydroquinone decreased the viability more markedly in cells expressing the G233E lipase variant, than in cells producing wild-type lipase. The more substantial viability loss was due to increased endoplasmic reticulum stress, as demonstrated by elevated levels of X-box binding protein 1 mRNA splicing and immunoglobulin binding protein, NAD(P)H:quinone oxidoreductase 1 and C/EBP homologous protein expression. Unresolved endoplasmic reticulum stress, and especially up-regulation of the pro-apoptotic transcription factor C/EBP homologous protein were likely responsible for the increased cell death. Our observations demonstrated that the combination of hydroquinone and misfolding pancreatic lipase variant promote increased levels of endoplasmic reticulum stress and cell death, which may predispose to chronic pancreatitis.
突变诱导的胰腺分泌酶蛋白错误折叠和随之而来的内质网应激可导致慢性胰腺炎。最近的一项研究表明,香烟烟雾也通过内质网应激增加了这种疾病的风险。在这里,我们使用哺乳动物细胞系研究了 G233E 错误折叠人胰腺脂肪酶变体和对苯二酚(香烟烟雾的主要有毒成分)的累积细胞效应。我们发现对苯二酚通过程序性细胞死亡以剂量依赖的方式降低细胞活力,并减少脂肪酶分泌,而不影响其表达。有趣的是,与产生野生型脂肪酶的细胞相比,对苯二酚在表达 G233E 脂肪酶变体的细胞中更显著地降低了细胞活力。存活能力的显著降低是由于内质网应激增加,这表现为 X 盒结合蛋白 1 mRNA 剪接和免疫球蛋白结合蛋白、NAD(P)H:醌氧化还原酶 1 和 C/EBP 同源蛋白表达水平升高。未解决的内质网应激,特别是促凋亡转录因子 C/EBP 同源蛋白的上调,可能是细胞死亡增加的原因。我们的观察表明,对苯二酚和错误折叠胰腺脂肪酶变体的组合促进了内质网应激和细胞死亡水平的增加,这可能导致慢性胰腺炎。