Department of Otolaryngology-Head and Neck, Jinling Hospital, Medical College of Nanjing University, Nanjing 210002, Jiangsu, China.
Department of Otolaryngology-Head and Neck, The First School of Clinical Medicine, Southern Medical University, Nanjing 210002, Jiangsu, China.
Aging (Albany NY). 2022 Oct 27;14(20):8486-8497. doi: 10.18632/aging.204356.
Excessive pepsin can damage both normal laryngeal epithelial cells and laryngeal cancer (LC) cells. Heat shock protein 70 (HSP70) is closely related to pepsin. In this paper, we will explore the different significance of the regulatory role of HSP70 in endoplasmic reticulum stress (ERS) level in pepsin-treated laryngeal epithelial cells and LC cells.
In cell experiments, laryngeal epithelial cells and LC cells were selected and induced by different concentrations of pepsin. Cell activity was detected by CCK8, cell apoptosis was detected by flow cytometry, and autophagy was detected by autophagy detection kit. The expression of ER)-related proteins was detected by immunofluorescence (IF) and Western blot. Cell transfection was used to inhibit HSP70 expression in both cells, and ERS, apoptosis, and autophagy were measured using related techniques. In animal experiments, a mouse model bearing LC was established. TUNEL assay detected apoptosis, autophagy kit detected autophagy, and ER-related protein expression was detected by Western blot.
HSP70 was increased in pepsin-stimulated laryngeal epithelial cells and LC cells, thereby inhibiting ER and ER-induced apoptosis and autophagy. Inhibition of HSP70 reduced the expression of glucose regulated protein 78 (GRP78) in pepsin-stimulated laryngeal epithelial cells and LC cells, and only inhibited downstream apoptosis-related pathways in laryngeal epithelial cells rather than in LC cells. Inhibition of HSP70 and ER could significantly promote apoptosis and inhibit tumor growth in the absence of pepsin stimulation .
ER level regulated by HSP70 had different significance in laryngeal epithelial cells and LC cells treated with pepsin.
过量的胃蛋白酶可损伤正常喉上皮细胞和喉癌（LC）细胞。热休克蛋白 70（HSP70）与胃蛋白酶密切相关。在本文中，我们将探讨 HSP70 在胃蛋白酶处理的喉上皮细胞和 LC 细胞中对内质网应激（ERS）水平的调节作用的不同意义。
在细胞实验中，选择喉上皮细胞和 LC 细胞，并分别用不同浓度的胃蛋白酶诱导。通过 CCK8 检测细胞活性，通过流式细胞术检测细胞凋亡，通过自噬检测试剂盒检测自噬。通过免疫荧光（IF）和 Western blot 检测 ER 相关蛋白的表达。用细胞转染抑制两种细胞中的 HSP70 表达，并使用相关技术测量 ERS、凋亡和自噬。在动物实验中，建立了 LC 荷瘤小鼠模型。TUNEL 检测凋亡，自噬试剂盒检测自噬，Western blot 检测 ER 相关蛋白的表达。
胃蛋白酶刺激的喉上皮细胞和 LC 细胞中 HSP70 增加，从而抑制 ER 和 ER 诱导的凋亡和自噬。抑制 HSP70 降低了胃蛋白酶刺激的喉上皮细胞和 LC 细胞中葡萄糖调节蛋白 78（GRP78）的表达，但仅抑制了喉上皮细胞而不是 LC 细胞中的下游凋亡相关途径。在没有胃蛋白酶刺激的情况下，抑制 HSP70 和 ER 可显著促进凋亡并抑制肿瘤生长。
在胃蛋白酶处理的喉上皮细胞和 LC 细胞中，HSP70 调节的 ER 水平具有不同的意义。
