3-溴丙酮酸增加MCF-7/TR耐药细胞对他莫昔芬的敏感性  

3-Bromopyruvic acid increases the sensitivity of MCF-7/TR cells to tamoxifen

作　　者：张璟钰 陈宗跃 刘婷婷[1] 张敏琴 酆啸 邹宇驰 沈祥春[1] 陈妍[1] ZHANG Jing-yu;CHEN Zong-yue;LIU Ting-ting;ZHANG Min-qin;FENG Xiao;ZOU Yu-chi;SHEN Xiang-chun;CHEN Yan(Key Laboratory of Optimal Utilization of Natural Medicine Resources,Guizhou Medical University,the High Educational Key Laboratory of Guizhou Province for Natural Medicinal Pharmacology and Druggability,Guiyang 550025,China)

机构地区：[1]贵州医科大学天然药物资源优效利用重点实验室,贵州省高等学校天然药物药理与成药性评价重点实验室,贵州贵阳550025

出　　处：《药学学报》2020年第10期2392-2397,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81860648);2016年度贵州省高层次创新型人才计划(71000305);贵州省普通高等学校科技拔尖人才(黔教合KY字[2018]048);贵州省2018年大学生创新创业训练计划项目(2018520337);贵州省高层次创新型人才百层次人才(贵州省科技厅黔科合人才[2015]4029号);贵州省科技计划项目(黔科合[2016]支撑2826);贵州省科技计划项目(黔科合平台人才[2017]5802号)。

摘　　要：本研究探讨3-溴丙酮酸增加人乳腺癌他莫昔芬耐药细胞MCF-7/TR对他莫昔芬敏感性的作用及机制。4-羟基他莫昔芬(4-OHT)是他莫昔芬在体内的活性形式,本研究采用4-OHT进行实验。MTT实验检测3-溴丙酮酸对MCF-7/TR细胞活力的影响,并通过显微镜观察细胞形态,选用对MCF-7/TR细胞的无毒剂量进行研究;检测4-OHT单用或联合3-溴丙酮酸处理MCF-7/TR耐药细胞的IC50,计算逆转耐药倍数;乳酸检测试剂盒检测MCF-7和MCF-7/TR细胞的乳酸水平;Western blot实验检测MCF-7和MCF-7/TR细胞中葡萄糖转运蛋白1(glucose transporter 1,GLUT1)、己糖激酶2(hexokinase 2,HK2)、乳酸脱氢酶A(lactate dehydrogenase A,LDHA)的表达水平。结果显示,40μmol·L-1的3-溴丙酮酸对MCF-7/TR细胞活力无明显影响;3-溴丙酮酸可显著增加4-OHT对MCF-7/TR耐药细胞的抑制作用,其逆转耐药倍数为1.91倍;MCF-7/TR耐药细胞中乳酸水平和GLUT1、HK2、LDHA蛋白的表达显著高于MCF-7细胞;与单用4-OHT相比,联用3-溴丙酮酸显著减少了MCF-7/TR耐药细胞中乳酸水平及GLUT1、HK2、LDHA蛋白的表达。以上研究结果表明,3-溴丙酮酸可增加MCF-7/TR耐药细胞对他莫昔芬的敏感性,其机制与抑制有氧糖酵解有关。We investigated the ability of 3-bromopyruvic acid to increase the sensitivity of tamoxifen-resistant MCF-7/TR cells to tamoxifen and to explore the underlying mechanism.4-Hydroxy tamoxifen(4-OHT)is the active form of tamoxifen in vivo and was used in this study.The effect of 3-bromopyruvic acid on the viability of MCF-7/TR cells was measured by MTT assay and the morphology of MCF-7/TR cells was observed with an inverted microscope.The results show that 3-bromopyruvic acid at 40μmol·L-1has no significant effect on the viability of MCF-7/TR cells,and this concentration was used in the subsequent experiments:3-bromopyruvic acid significantly increased the inhibitory effect of 4-OHT on the viability of MCF-7/TR cells,with a reversal-fold of 1.91,as determined by MTT assay;the lactate level,determined with a lactate detection kit,and the expression levels of GLUT1,HK2,and LDHA in MCF-7/TR cells as measured by immunoblotting were significantly higher than in MCF-7 cells;and compared with 4-OHT treatment alone,the combination treatment of 3-bromopyruvic acid and4-OHT significantly reduced the lactate level and the expression of GLUT1,HK2,LDHA in MCF-7/TR cells.The above results show that 3-bromopyruvic acid increases the sensitivity of MCF-7/TR cells to tamoxifen,and the mechanism is related to the inhibition of aerobic glycolysis.

关 键 词：乳腺癌 耐药 他莫昔芬 3-溴丙酮酸 有氧糖酵解 

分 类 号：R737.9[医药卫生—肿瘤]