G蛋白偶联雌激素受体抑制剂增加他莫昔芬诱导T-47DTR耐药细胞凋亡  

GPER inhibitor increases tamoxifen-induced apoptosis in T-47DTR-resistant cells

作　　者：张敏琴 宋雨萱 樊双琴 任爽 张玥[2] 陈妍[2] 沈祥春[2] 刘同征 张敏 无[3] ZHANG Min-qin;SONG Yu-xuan;FAN Shuang-qin;REN Shuang;ZHANG Yue;CHEN Yan;SHEN Xiang-chun;LIU Tong-zheng;ZHANG Min;无(Dept of Physiology,School of Basic Medicine;Key Laboratory of Optimal Utilization of Natural Medicinal Resources,Guizhou Medical University,Guiyang 550025,China;Jinan University Institute of Tumor Pharmacology Research,Guangzhou 510632,China)

机构地区：[1]贵州医科大学基础医学院生理学教研室,贵州贵阳550025 [2]贵州医科大学天然药物资源优效利用重点实验室,贵州贵阳550025 [3]暨南大学肿瘤药理研究所,广东广州510632

出　　处：《中国药理学通报》2023年第1期96-100,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81860648);贵州省普通高等学校科技拔尖人才支持计划(黔教合KY字[2018]048);贵州省药学国际科技合作基地(黔科合平台人才[2017]5802);贵州省优秀青年科技人才计划项目(黔科合平台人才[2021]5632号);贵州省普通高等学校青年科技人才成长项目(黔教合KY字[2021]169);贵州省科技计划项目(黔科合基础-ZK[2021]一般527)。

摘　　要：目的研究G蛋白偶联雌激素受体(G-protein coupled estrogen receptor,GPER)抑制剂G15对乳腺癌他莫昔芬耐药细胞T-47DTR敏感性的影响。方法采用他莫昔芬在体内活性形式4-羟基他莫昔芬(4-OHT)进行实验。MTT实验检测乳腺癌他莫昔芬耐药细胞系T-47DTR及其亲本细胞系T-47D对他莫昔芬的敏感性;膜浆分离分析抑制剂G15对GPER蛋白表达的变化;流式细胞术AnnexinV-FITC/PI双染分析4-OHT联用G15对T-47DTR细胞凋亡的影响;Western blot分析凋亡相关蛋白Bax、Bcl-2、caspase-3、cleaved-caspase-3、caspase-9、cleaved-caspase-9的表达水平。结果(1)相比于亲本细胞T-47D,T-47DTR耐药细胞对4-OHT的抵抗性显著增强。(2)给予4-OHT(2μmol·L^(-1)),与对照组相比,GPER在细胞膜分布增加,表明在T-47DTR耐药细胞中GPER被激活;与单用4-OHT相比,联用G15(5μmol·L^(-1))显著减少GPER表达。(3)GPER抑制剂G15能增加T-47DTR耐药细胞凋亡率同时下调抗凋亡蛋白Bcl-2,上调促凋亡蛋白Bax、cleaved caspase-3、cleaved caspase-9的表达。结论GPER抑制剂G15增加T-47DTR细胞的凋亡恢复耐药细胞对他莫昔芬的敏感性。Aim To study the effect of G protein-coupled estrogen receptor(GPER)inhibitor G15 on the sensitivity of breast cancer tamoxifen-resistant cells to T-47DTR.Methods Experiments were carried out with 4-hydroxytamoxifen(4-OHT),the active form of tamoxifen in vivo.The sensitivity of tamoxifen-resistant breast cancer cell line T-47DTR and its parental cell line T-47D to tamoxifen was detected by MTT assay;the expression of GPER protein was analyzed by plasma separation of inhibitor G15;the effect of 4-OHT combined with G15 on the apoptosis of T-47DTR cells was analyzed by flow cytometry AnnexinV-FITC/PI double staining;the expression levels of apoptosis-related proteins Bax,Bcl-2,caspase-3,cleaved caspase-3,caspase-9,cleaved caspase-9 were analysed by Western blot.Results(1)Compared with the parental cell T-47D,the resistance of T-47DTR-resistant cells to 4-OHT was significantly enhanced.(2)When 4-OHT(2μmol·L^(-1))was administered,the membrane distribution of GPER increased,indicating that GPER was activated in T-47DTR-resistant cells compared with the control group;Compared with OHT,the use of G15(5μmol·L^(-1))and OHT significantly reduced the expression of GPER.(3)GPER inhibitor G15 could increase the apoptotic rate of T-47DTR-resistant cells while down-regulating the anti-apoptotic protein Bcl-2 and up-regulating the expression of pro-apoptotic proteins Bax,cleaved caspase-3,cleaved caspase-9.Conclusions The GPER inhibitor G15 increases the apoptosis of T-47DTR cells and restores the sensitivity of drug-resistant cells to tamoxifen.

关 键 词：乳腺癌 他莫昔芬 耐药 GPER G15 凋亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R392.11[医药卫生—基础医学] R737.9R977.12R979.1