Effect and mechanisms of celastrol on the apoptosis of HOS osteosarcoma cells
Effect and mechanisms of celastrol on the apoptosis of HOS osteosarcoma cells
复制标题
雷公藤红素对HOS骨肉瘤细胞凋亡的影响及机制
DOI：
10.3892/or.2018.6619
复制
发表时间：
2018-10-01
期刊：
影响因子：
4.2
通讯作者：
He, Bin
中科院分区：
文献类型：
--
作者：
Chen, Yanyang;Ou, Yunsheng;He, Bin
骨肉瘤是最常见的骨原发恶性肿瘤，多见于儿童和青少年，可导致早期转移和预后不良。本研究主要探讨雷公藤红素对骨肉瘤HOS细胞凋亡和自噬的影响及其相关机制。用细胞计数试剂盒8、流式细胞仪、倒置相差显微镜、Hoechst染色和透射电子显微镜检测细胞活力、细胞内钙离子浓度、细胞凋亡率和细胞形态的变化。免疫印迹法检测内质网应激相关蛋白(BiP、PERK、P-PERK、IRE1α、Calnexin、Pdi和Erol-Lα)、凋亡相关蛋白(CHOP、裂解caspase-12)、线粒体凋亡相关蛋白(Bax、Bcl2和细胞色素c)、裂解Caspase-3、自噬相关蛋白(Lc3-I、Lc3-II和P62)和β-肌动蛋白的表达水平。雷公藤红素显著抑制HOS细胞活性，并呈剂量依赖关系，促进ERS相关蛋白、凋亡相关蛋白和线粒体凋亡相关蛋白的表达。ERS抑制剂牛磺酸脱氧胆酸可促进雷公藤红素诱导的HOS细胞自噬和凋亡。预先给予PERK抑制剂GSK2656157可显著促进雷公藤红素诱导的死亡和抑制HOS细胞的自噬。提示ERS途径和线粒体途径参与了雷公藤红素诱导HOS细胞的凋亡。ERS/PERK通路可能对雷公藤红素诱导的HOS细胞凋亡具有保护作用，并可能在自噬过程中发挥复杂的作用。
Osteosarcoma is the most common primary malignant tumor of the bone found predominantly in children and teenagers and results in early metastasis and poor prognosis. The present study primarily focused on the impact of celastrol on apoptosis and autophagy of osteosarcoma HOS cells, as well as the related mechanisms. Following the appropriate treatment, the human osteosarcoma cell line HOS was assessed for viability, Ca2+ in cells, apoptosis and changes in cell morphology using Cell Counting Kit-8, flow cytometry, inverted phase contrast microscope, Hoechst staining and transmission electron microscopy. The expression levels of various proteins, including endoplasmic reticulum stress (ERS)-related proteins (Bip, PERK, p-PERK, IRE1 alpha, calnexin, PDI and Erol-L alpha), apoptosis-related proteins (CHOP, cleaved caspase-12), mitochondrial apoptosis-related proteins (Bax, Bcl-2 and cytochrome c), cleaved caspase-3, and autophagy-related proteins (LC3-I, LC3-II and P62) and beta-actin, were assessed with western blotting. Celastrol significantly inhibited the viability of HOS cells in a dose-dependent manner and promoted the expression of ERS-related, apoptosis-related and mitochondrial apoptosis-related proteins. The ERS inhibitor tauroursodeoxycholate promoted celastrol-induced autophagy and apoptosis of HOS cells. Pretreatment with the PERK inhibitor GSK2656157 significantly promoted celastrol-induced death and attenuated HOS cell autophagy. Our results indicated that the ERS pathway and the mitochondrial pathway were involved in celastrol-induced apoptosis of HOS cells. The ERS/PERK pathway may protect HOS cells from apoptosis by celastrol and may play a complicated role in the process of autophagy.