木香烃内酯通过内质网应激和自噬诱导骨肉瘤细胞凋亡  

Costunolide induces apoptosis of osteosarcoma cells through endoplasmic reticulum stress and autophagy

作　　者：刘景新 陈余兴 王贵 LIU Jingxin;CHEN Yuxing;WANG Gui(Department of Orthopedics,Hainan Western Central Hospital,Danzhou 571700,China)

机构地区：[1]海南西部中心医院骨科,儋州571700

出　　处：《中国免疫学杂志》2022年第15期1803-1807,1812,共6页Chinese Journal of Immunology

基　　金：海南省卫生健康行业科研项目(20A200368)资助。

摘　　要：目的:探讨木香烃内酯(Cos)对骨肉瘤(OS)细胞凋亡的影响及其相关分子机制。方法:体外培养MG63细胞,采用不同浓度(5、10、15、20、25、40μmol/L)Cos处理MG63细胞48 h。CCK-8法检测细胞增殖抑制率,划痕试验检测细胞迁移能力,Transwell小室法测定细胞侵袭能力,Annexin V-FITC/PI双染法检测细胞凋亡率,Western blot法检测B细胞淋巴瘤-2(Bcl-2)、Bcl-2结合抗凋亡基因(Bax)、Caspase-3、Cleaved Caspase-3、真核生物翻译起始因子2α(eIF2α)、C/EBP同源蛋白(CHOP)、蛋白激酶R样内质网激酶(PERK)及微管相关蛋白轻链3B-Ⅱ(LC3B-Ⅱ)、Beclin-1、p62蛋白表达水平。结果:Cos显著降低MG36细胞增殖活力,且其抑制作用呈浓度和时间依赖性(P<0.05)。Cos显著降低MG36细胞迁移、侵袭能力,提高细胞凋亡率,差异具有统计学意义(P<0.05)。Cos可显著上调MG36细胞的eIF2α、CHOP、PERK蛋白表达水平,上调MG36细胞中LC3B-Ⅱ、Beclin-1蛋白表达水平,下调p62蛋白表达水平,差异均有统计学意义(P<0.05)。Cos显著下调Bcl-2蛋白表达水平,上调Bax、Caspase-3、Cleaved Caspase-3蛋白表达水平,差异具有统计学意义(P<0.05);内质网应激抑制剂牛磺熊去氧胆酸(TUDC)和自噬抑制剂3-甲基腺嘌呤(3-MA)均可逆转Cos对MG36细胞凋亡相关蛋白的调控作用。结论:Cos通过介导OS细胞内质网应激和自噬促进OS细胞凋亡。Objective:To investigate the effect of costunolide(Cos)on apoptosis of osteosarcoma(OS)cells and its related molecular mechanism.Methods:MG63 cells were cultured in vitro and treated with different concentrations of Cos(5,10,15,20,25,40 μmol/L)for 48 h. Cell proliferation inhibition rate was detected by CCK-8 method,cell migration ability was detected by scratch test,cell invasive ability was detected by Transwell chamber method,cell apoptosis rate was detected by Annexin V-FITC/PI double staining method,the expressions of B cell lymphoma 2(Bcl-2),Bcl-2 binding anti apoptosis gene(Bax),Caspase-3,Cleaved Caspase-3,eukaryotic translation initiation factor 2α(eIF2α),C/EBP homologous protein(CHOP),protein kinase R like endoplasmic reticulum kinase(PERK),microtubule-associated protein light chain 3B-Ⅱ(LC3B-Ⅱ),Beclin-1 and P62 were detected by Western blot.Results:Cos significantly decreased the proliferation activity of MG36 cells in a concentration and time-dependent manner(P<0.05). Cos significantly reduced the migration and invasion ability of MG36 cells and increased the apoptosis rate(P<0.05). Cos significantly up-regulated the expression levels of eIF2α,CHOP and PERK in MG36 cells,up-regulated the expression levels of LC3B-Ⅱ and Beclin-1 in MG36 cells,and down-regulated the expression level of p62 protein(P<0.05). Cos significantly down-regulated the expression of Bcl-2 and up-regulated the expressions of Bax,Caspase-3 and Cleaved Caspase-3(P<0.05);endoplasmic reticulum stress inhibitor tauroursodeoxycholic acid(TUDC)and autophagy inhibitor 3-methyladenine(3-MA)could reverse the regulation of apoptosis related proteins by Cos in MG36 cells.Conclusion:Cos promotes OS cell apoptosis by mediating endoplasmic reticulum stress and autophagy.

关 键 词：木香烃内酯 骨肉瘤 凋亡 自噬 内质网应激 

分 类 号：R738.1[医药卫生—肿瘤]