冬凌草甲素通过抑制STAT-3/SYNPO-2信号通路诱导人恶性胶质瘤U87MG细胞凋亡  

Oridonin promotes apoptosis of human malignant glioma U87MG cells via STAT-3/SYNPO-2 signaling pathway

作　　者：陈爽[1] 王宸 马晓娇 邱红梅[1] CHEN Shuang;WANG Chen;MA Xiao-jiao;QIU Hong-mei(Department of Pharmacology,Chongqing Key Lab of Biochemistry and Molecular Pharmacology,Chongqing Key Research Laboratory for Drug Metabolism,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学药理学教研室,重庆市生物化学与分子药理学重点实验室,药物代谢研究重庆市重点实验室,重庆400016

出　　处：《中国病理生理杂志》2021年第10期1801-1807,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31400881);重庆市自然科学基金资助项目(No.cstc2017jcyjAX0211);重庆医科大学药学院科研资助项目(No.YXY2019SDTR01)。

摘　　要：目的:研究冬凌草甲素(ORI)在体外对人源恶性胶质瘤U87MG细胞的作用,并探讨其机制是否涉及信号转导及转录激活因子3(STAT-3)/突触足蛋白2(SYNPO-2)信号通路。方法:体外培养U87MG细胞,采用不同浓度(2.5、5、7.5、10和12.5μmol/L)的ORI处理后,CCK-8法检测U87MG细胞活力;划痕实验检测U87MG细胞的迁移能力;流式细胞术检测U87MG细胞的凋亡;给予STAT-3的磷酸化激动剂白细胞介素6(IL-6)和抑制剂Stattic进行干预,将细胞分为对照(control)组、ORI组、IL-6组、Stattic组、IL-6+ORI组和Stattic+ORI组,Western blot检测Bcl-2、Bax、STAT-3、p-STAT-3(Tyr705)和SYNPO-2蛋白水平。结果:ORI呈浓度和时间依赖性地显著抑制U87MG细胞的活力和迁移能力(P<0.01),并促进其凋亡(P<0.01);Western blot结果显示,ORI可上调Bax并下调Bcl-2蛋白的表达(P<0.01),且使p-STAT-3(Tyr705)蛋白水平显著降低(P<0.01),SYNPO-2蛋白表达显著增高(P<0.01),而STAT-3蛋白含量基本不变;应用Stattic可协同促进ORI的促凋亡作用(P<0.01),而IL-6则逆转ORI的作用(P<0.01)。结论:体外细胞实验中,ORI可显著抑制恶性胶质瘤U87MG细胞的活力和迁移能力,并促进其凋亡,其作用机制可能与抑制STAT-3/SYNPO-2信号通路有关。AIM:To investigate the effect of oridonin(ORI)on human malignant glioma U87 MG cells in vitro,and to explore whether the mechanism is involved in the inhibition of signal transducer and activator of transcription-3(STAT-3)/synaptopodin-2(SYNPO-2)signaling pathway.METHODS:The U87 MG cells were cultured in vitro and treated with ORI at different concentrations(2. 5,5,7. 5,10 and 12. 5 μmol/L),and then the viability of U87 MG cells was measured by CCK-8 assay. The migration of U87 MG cells was detected by scratch test. The apoptosis of U87 MG cells was measured by flow cytometry. The STAT-3 phosphorylation agonist interleukin-6(IL-6)and inhibitor Stattic were given for intervention,and the U87 MG cells were divided into control group,ORI group,IL-6 group,Stattic group,IL-6+ORI group and Stattic+ORI group. The protein levels of Bcl-2,Bax,STAT-3,p-STAT-3(Tyr705)and SYNPO-2 were determined by Western blot.RESULTS:Treatment with ORI significantly inhibited the viability and migration of U87 MG cells(P<0. 01),and promoted their apoptosis(P<0. 01)in a concentration-and time-dependent manner. Western blot analysis showed that ORI up-regulated the protein expression of Bax but down-regulated the protein expression of Bcl-2,and the protein level of p-STAT-3(Tyr705)was significantly decreased(P<0. 01),the protein expression of SYNPO-2 was significantly increased(P<0. 01),while STAT-3 protein was unobviously changed. The application of Stattic synergistically promoted the pro-apoptotic effect of ORI(P<0. 01),while IL-6 reversed this effect(P<0. 01).CONCLUSION:Treatment with ORI significantly inhibits the viability and migration of malignant glioma U87 MG cells in vitro,and promotes their apoptosis. The mechanism is perhaps involved in the inhibition of STAT-3/SYNPO-2 signaling pathway.

关 键 词：冬凌草甲素 胶质瘤 细胞凋亡 STAT-3/SYNPO-2信号通路 

分 类 号：R739.4[医药卫生—肿瘤] R363.2[医药卫生—临床医学]