人参皂苷辛酸酯通过内质网应激诱导肝癌细胞免疫原性细胞死亡的机制研究
Mechanisms of immunogenic cell death induced by octyl ester derivative of ginsenoside Rh2 in hepatocellular carcinoma cells based on endoplasmic reticulum stress
戴珍珍 1黄青昕 1胡淇睿 2吴汉城 1潘瑶 1邓泽元 2陈芳1
作者信息
- 1. 南昌大学公共卫生学院,南昌 330006
- 2. 食品科学与资源挖掘全国重点实验室,南昌大学,南昌 330047
- 折叠
摘要
目的:探讨人参皂苷辛酸酯(Rh2-O)能否诱导Huh-7肝癌细胞免疫原性细胞死亡及可能机制.方法:体外培养Huh-7细胞,设置空白对照组、Rh2-O药物组及阳性对照组(米托蒽醌处理),CCK-8、流式细胞术分别检测细胞活力和凋亡,ELISA及化学发光法分别检测细胞上清高迁移率族蛋白1(HMGB1)和三磷酸腺苷(ATP)含量,免疫荧光法检测细胞钙网蛋白(CRT)的膜外翻,荧光探针DCFH-DA检测细胞内ROS水平,Western blot检测细胞内质网应激信号通路相关蛋白表达.结果:Rh2-O可显著降低细胞活力,促进细胞凋亡,诱导细胞HMGB1分泌、ATP释放及CRT膜外翻,使细胞内蓄积大量ROS,提高内质网应激信号通路相关蛋白PERK、eIF2α、p-eIF2α表达(均P<0.05).内质网应激抑制剂4-苯基丁酸(4-PBA)预处理可显著减弱Rh2-O诱导的CRT膜外翻(P<0.05).结论:Rh2-O可诱导肝癌细胞发生免疫原性细胞死亡,其机制可能与Rh2-O激活内质网应激促进细胞CRT膜外翻有关.
Abstract
Objective:To investigate whether octyl ester derivative of ginsenoside Rh2(Rh2-O)can induce immunogenic cell death of Huh-7 hepatocellular carcinoma cells and possible mechanism.Methods:Huh-7 cells were cultured in vitro,and divided into control group,Rh2-O group,positive control group(mitoxantrone treatment).Viability and apoptosis of cells were detected by CCK-8 and flow cytometry,respectively.Concentrations of high mobility family protein 1(HMGB1)and adenosine triphosphate(ATP)in supernatant were detected by ELISA and chemiluminescence assay,respectively.Membrane eversion of calreticulin(CRT)was detected by immunofluorescence assay.ROS level in cells was detected by fluorescence probe DCFH-DA,and expressions of proteins associated with endoplasmic reticulum stress signaling pathway were detected by Western blot.Results:Rh2-O treatment significantly reduced cell viability,promoted apoptosis,induced secretion of HMGB1,ATP,membrane eversion of CRT,increased accumulation of ROS in cells,and enhanced expressions of endoplasmic reticulum stress-related proteins PERK,eIF2α,p-eIF2α(all P<0.05).After addition of endoplasmic reticulum stress inhibitor 4-phenylbutyric acid(4-PBA),membrane eversion of CRT induced by Rh2-O was significantly inhibited(P<0.05).Conclusion:Rh2-O can induce immunogenic cell death in hepatocellular carcinoma cells,whose mechanism may be associated with activation of endoplasmic reticulum stress and promotion of CRT membrane eversion.
关键词
人参皂苷/辛酸酯衍生物/肝癌/免疫原性细胞死亡/内质网应激Key words
Ginsenoside Rh2/Octyl ester derivatives/Hepatocellular carcinoma/Immunogenic cell death/Endoplasmic reticulum stress引用本文复制引用
基金项目
国家自然科学基金(81860578)
江西省主要学科学术和技术带头人培养计划-青年人才项目(20204BCJ23025)
江西省研究生创新专项资金项目(YC2021-S075)
出版年
2024
国家科技期刊平台
NSTL
万方数据