雷公藤红素通过内质网应激相关通路促进人骨肉瘤HOS细胞凋亡

发布时间：2018-05-03 18:56

本文选题：骨肉瘤 + 内质网应激　；参考：《肿瘤》2017年09期

【摘要】：目的:观察雷公藤红素对人骨肉瘤HOS细胞凋亡的影响,并探讨其可能的作用机制。方法 :用不同浓度(0、1.5、2.5、3.5和4.5μmol/L)的雷公藤红素处理骨肉瘤HOS细胞24 h后,采用CCK-8法检测细胞活性,确定最佳实验浓度。采用倒置相差显微镜观察细胞形态变化,Hoechst33258染色后相差显微镜观察细胞凋亡的形态学改变,FCM法检测细胞凋亡率和细胞内Ca2+浓度变化,蛋白质印迹法检测结合免疫球蛋白(binding immunoglobulin protein,BIP)、钙联蛋白(calnexin,CNX)、内质网氧化还原酶1-Lα(endoplasmic reticulum oxidoreductin 1-L alpha,Ero1-Lα)、蛋白质二硫键异构酶(protein disulfide isomerase,PDI)、肌醇依赖酶1(αinositol-requiring enzyme 1 alpha,IRE1α)、蛋白激酶样内质网激酶(protein kinase-like endoplasmic reticulum kinase,PERK)和磷酸化PERK(phosphorylatedPERK,p-PERK)、CCAAT/增强子结合蛋白同源蛋白(CCAAT/enhancerbinding protein homologous protein,CHOP)、剪切型caspase-12(cleavedcaspase-12,c-caspase-12)和c-caspase-3的表达变化。结果 :不同浓度的雷公藤红素均可抑制HOS细胞活性(P值均0.05),且呈浓度依赖性。3μmol/L雷公藤红素处理24 h后,HOS细胞形态明显萎缩、紊乱,漂浮细胞增多,且出现明显核固缩和核碎裂等典型的细胞凋亡形态学改变。与未处理对照组相比,3μmol/L雷公藤红素处理24 h后细胞凋亡率和细胞内Ca2+浓度明显升高(P值均0.05)。3μmol/L雷公藤红素处理24 h后,内质网应激相关标志蛋白BIP、CNX、Ero1-Lα、PDI、IRE1α和p-PERK以及内质网应激介导凋亡相关蛋白CHOP、c-caspase-12和c-caspase-3的表达水平明显升高(P值均0.05)。结论 :雷公藤红素可诱导人骨肉瘤HOS细胞凋亡,其作用机制可能与内质网应激介导细胞凋亡通路有关。
[Abstract]:Aim: to investigate the effect of tripterine on apoptosis of human osteosarcoma HOS cells and its possible mechanism. Methods: osteosarcoma HOS cells were treated with tripterine at different concentrations of 1.5 渭 mol / L and 4.5 渭 mol / L for 24 h. The cell activity was detected by CCK-8 assay and the optimal concentration was determined. Morphological changes of apoptosis were observed by inverted phase contrast microscope (PPM) and Hoechst33258 staining. The apoptosis rate and intracellular Ca2 concentration were detected by FCM. Western blot detection of binding immunoglobulin, calnexin immunoglobulin, endoplasmic redoxoreductase 1-L 伪 -endoplasmic reticulum oxidoreductin 1-L alpha-Ero1-L 伪, protein disulfide isomerase, inositol dependent enzyme 1 (伪 inositol-requiring enzyme 1 alpha-IRE1 伪, protein kinase-like endoplasmic reticulum kinase kinase-like endoplasmic) The expression of CCAATA / enhancer binding protein, CCAATR / Enhancer-binding protein homologous protein, Caspase-12 caspase-12cleavedcaspase-12c-caspase-12) and c-caspase-3 were detected by reticulum kinase- PERK) and phosphorylated PERKN (phosphorylated PERKA) and phosphorylated PERKN (phosphorylated PERKN). Results: Tripterygium wilfordii inhibited the activity of HOS cells in a dose-dependent manner (P < 0.05). After treated with tripterine for 24 h, the morphology of Hos cells shrank significantly, and the number of floating cells increased. Typical morphological changes of apoptosis such as nuclear pyknosis and nuclear fragmentation were also observed. Compared with the untreated control group, the apoptosis rate and intracellular Ca2 concentration of tripterine treated with 3 渭 mol/L tripterine for 24 h increased significantly (P = 0.050.30 渭 mol/L) after treatment with tripterine for 24 h. The expression of endoplasmic reticulum stress-related marker protein BIP-CNX1-L 伪, PDII-IRE1 伪 and endoplasmic reticulum stress-mediated apoptosis-associated protein CHOP-c-caspase-12 and c-caspase-3 increased significantly (P < 0.05). Conclusion: Tripterygium wilfordii can induce apoptosis of human osteosarcoma HOS cells and its mechanism may be related to endoplasmic reticulum stress-mediated apoptosis pathway.
【作者单位】：重庆医科大学附属第一医院骨科;
【基金】：国家自然科学基金资助项目(编号:81572634) 重庆市教育委员会研究生科研创新项目(编号:CYS15141)~~
【分类号】：R738.1

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