吉林大学学报（医学版）2025，Vol.51Issue(2)：420-427,8.DOI:10.13481/j.1671-587X.20250216
胡桃醌对骨肉瘤细胞凋亡和焦亡的影响
Effect of Juglone on apoptosis and pyroptosis of osteosarcoma cells
摘要
Abstract
Objective:To investigate the effects of Juglone on the apoptosis of osteosarcoma(OS)cells(U2OS and MG63 cells)through the cysteinyl aspartate specific proteinase-3(Caspase-3)/gasdermin E(GSDME)-mediated pyroptosis pathway.Methods:The U2OS and MG63 cells were cultured in vitro and divided into control group,different concentrations(5,10 and 20 μmol·L-1)of Juglone groups and Caspase-3 inhibitor Z-DEVD-FMK group(10 μmol·L-1 Juglone+30 μmol·L-1 Z-DEVD-FMK).The survival rates of cells in various groups were assessed by cell counting kit-8(CCK-8)assay,and the apoptotic rates were detected by flow cytometry.Lactate dehydrogenase(LDH)release assay was used to measure the release rates of LDH from the cells.Western blotting method was used to detect the expression levels of apoptosis-related proteins including B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),cleaved-Caspase-3 and poly(ADP-ribose)polymerase(PARP)and pyroptosis-related proteins including GSDME full form(GSDME-F)and GSDME N-terminal(GSDME-N).The levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)in the cell supernataut in various groups were measured by enzyme-linked immunosorbent assay(ELISA)method.Results:Compared with control group,the survival rates of cells in 5,10,and 20 μmol·L-1Juglone groups were significantly decreased(P＜0.05 or P＜0.01),and the 50％inhibitory concentration(IC50)values of U2OS cells and MG63 cells were 8.4 and 10.2 μmol·L-1,respectively.Compared with control group,the apoptotic rates and LDH release rates of U2OS and MG63 cells in 5 and 10 μmol·L-1Juglone groups were significantly increased(P＜0.05 or P＜0.01).Compared with control group,the expression levels of Bax,cleaved-Caspase-3,and cleaved-PARP proteins in 5 and 10 μmol·L-1 Juglone groups were significantly increased(P＜0.01),while the expression levels of Bcl-2 protein were significantly decreased(P＜0.01).Compared with control group,the levels of IL-1β and IL-18 in cell supernatant in 5 and 10 μmol·L-1Juglone groups were increased(P＜0.01).Compared with control group,the expression levels of cleaved-Caspase-3 and GSDME-N proteins in 5 and 10 μmol·L-1 Juglone groups were significantly increased(P＜0.01),while there was no difference in the expression level of GSDME-F protein(P＞0.05).Compared with 10 μmol·L-1 Juglone group,the expression levels of cleaved-Caspase-3 and GSDME-N in Z-DEVD-FMK group were significantly decreased(P＜0.01),while there was no difference in the expression level of GSDME-F protein(P＞0.05).Conclusion:Juglone can induce the apoptosis of U2OS and MG63 cells and cause the Caspase-3/GSDME-mediated pyroptosis.关键词
胡桃醌/骨肉瘤/细胞凋亡/细胞焦亡/B细胞淋巴瘤2/B细胞淋巴瘤2相关X蛋白Key words
Juglone/Osteosarcoma/Apoptosis/Cell pyroptosis/B-cell lymphoma-2/B-cell lymphoma-2-associated X protein分类
医药卫生引用本文复制引用
赵杰瑞,籍铭辛,张钰涵,陈姝彤,郭玉苗,张巍,彭鹏..胡桃醌对骨肉瘤细胞凋亡和焦亡的影响[J].吉林大学学报（医学版）,2025,51(2):420-427,8.基金项目
广东省卫健委医学科学技术研究基金资助项目(A2021004) （A2021004）
吉林省科技厅科技发展计划项目(YDZJ202301ZYTS111) （YDZJ202301ZYTS111）
吉林省教育厅"十三五"科学技术研究项目(JJKH20220464KJ) （JJKH20220464KJ）
