二甲双胍与二氯乙酸盐对MYCN扩增神经母细胞瘤BE-2C细胞的协同抗肿瘤作用  

Synergistic anti-tumor effect of metformin and dichloroacetate in MYCN-amplified neuroblastoma BE-2C cells

作　　者：王文斌 任飞桦 胡少洋 刘欢 廖清船 陈红霞 任平 WANG Wen-bin;REN Fei-hua;HU Shao-Yang;LIU Huan;LIAO Qing-chuan;CHEN Hong-xia;REN Ping(Dept of Thyroid and Breast Surgery, Xianning Central Hospital, Xianning, Hubei 437100,China;School of Pharmacy, Hubei University of Science and Technology, Xianning, Hubei 437100,China;School of Public Health, Guizhou Medical University, Guiyang 550025,China)

机构地区：[1]咸宁市中心医院甲乳外科,湖北咸宁437100 [2]湖北科技学院药学院,湖北咸宁437100 [3]贵州医科大学公共卫生学院,贵州贵阳550025

出　　处：《中国药理学通报》2019年第9期1256-1261,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81502635);地方高校国家级大学生创新创业训练计划项目(No 201710927012);湖北省卫生健康委科研资助项目(No WJ2019M096);湖北科技学院药学重点专项科研项目(No 2018-19XZY12)

摘　　要：目的研究二甲双胍和二氯乙酸盐(dichloroacetate,DCA)单独与联合应用,对MYCN扩增神经母细胞瘤BE-2C细胞增殖的抑制作用及其作用机制。方法CCK-8法检测二甲双胍与DCA单独或联用对BE-2C细胞增殖的抑制作用;采用葡萄糖和乳酸测定试剂盒,检测葡萄糖吸收量和乳酸生成量的变化;Annexin V-FITC/PI双染法流式细胞术检测细胞凋亡;Western blot分析凋亡相关蛋白的表达变化。结果二甲双胍和DCA对BE-2C细胞增殖均具有明显的抑制作用,联用组的葡萄糖消耗和乳酸的产生量较二甲双胍单独用药组明显减少(P<0.01);联用组的细胞凋亡率较单独用药组均明显升高(P<0.01);联用组细胞的Bax、cleaved caspase-3蛋白表达水平较单用组均明显增加,而Bcl-2蛋白表达水平较单用组明显降低。结论二甲双胍联用DCA对MYCN扩增神经母细胞瘤BE-2C细胞具有协同抗肿瘤作用,且与减少二甲双胍引起的乳酸堆积和诱导肿瘤细胞发生凋亡有关。Aim To investigate the effect of metformin combined with dichloroacetate on the proliferation of MYCN -amplified neuroblastoma BE-2C cells and its mechanisms. Methods The inhibitory effects of metformin and dichloroacetate alone or in combination on BE-2C cells was measured by CCK-8 assay;the concentrations of glucose and lactic acid in the medium were measured by a glucose test kit and L-lactic acid assay kit;cell apoptosis was determined by flow cytometry (FCM) with Annexin V-FITC conjugated propidium iodide (PI) staining;the expressions of apoptosis related protein were detected by Western blot. Results Metformin and dichloroacetate showed significant proliferation inhibition activity on BE-2C cells;there was obviously decreased glucose uptake and lactic acid production of combined group compared with alone group( P <0.01);the apoptotic rate was significantly higher in combined group compared with that in alone group ( P <0.01);Bax and cleaved caspasce-3 protein expression markedly increased in combined group, but Bcl-2 protein expression significantly increased compared with alone group ( P <0.01). Conclusions Metformin combined with dichloroacetate shows a significant synergistic anti-tumor effect against BE-2C cells by reducing the accumulation of lactic acid caused by metformin and inducing apoptosis.

关 键 词：MYCN 扩增神经母细胞瘤 二甲双胍 二氯乙酸盐 协同抗肿瘤作用 乳酸堆积 细胞凋亡 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R329[医药卫生—基础医学]