摘要
目的 研究二萜衍生物Jar-TTA对糖酵解/氧化磷酸化(OXPHOS)的双重抑制作用,并探讨其抗食管癌机制。方法 MTT法检测Jar-TTA对食管癌细胞EC109、KYSE-150的抗增殖作用;荧光显微镜观察线粒体膜电位(MMP);流式细胞术定量分析细胞凋亡、MMP及葡萄糖摄取;能量代谢分析仪实时检测细胞糖酵解及OXPHOS;Western blot检测蛋白表达。结果 Jar-TTA明显抑制食管癌细胞的增殖,且呈浓度依赖性;2、4、8 μmol·L^-1 Jar-TTA诱导EC109早期凋亡率分别为(27.9±6.1)%、(71.1±9.3)%和(65.0±9.5)%,与对照组相比,差异均具有显著性( P <0.01);Jar-TTA明显抑制细胞的糖酵解和OXPHOS;此外,Jar-TTA诱导MMP下降,抑制葡萄糖摄取,同时下调葡萄糖转运蛋白4(GLUT4)及乳酸脱氢酶A(LDHA)表达。结论 Jar-TTA通过双重抑制糖酵解/OXPHOS诱导食管癌细胞凋亡,其机制可能与Jar-TTA降低MMP而干扰线粒体OXPHOS功能,以及下调GLUT4和LDHA而抑制葡萄糖摄取和糖酵解进程有关。
Aim To investigate the effects of Jar-TTA in dual inhibition of glycolysis/oxidative phosphorylation in human esophageal cancer cells and explore the related molecular mechanism.Methods The effect of Jar-TTA on the esophageal cancer cell EC109 and KYSE-150 viability was examined using MTT assay.The effect of Jar-TTA in apoptosis morphology and mitochondrial membrane potential(MMP) was observed by a fluorescence microscopy.The apoptosis of cell lines treated with Jar-TTA, the quantitative analysis of MMP falling, as well as the glucose uptake was analyzed by flow cytometry.The mitochondrial OXPHOS and glycolysis of EC109 cells in response to Jar-TTA were analyzed using a Seahorse XFp extracellular flux analyzer by real-time measurements of the oxygen consumption rate(OCR, indicative of mitochondrial OXPHOS) and extracellular acidification rate(ECAR, indicative of glycolysis).The expression of the proteins related with glycolysis were detected by Western blot.Results Jar-TTA caused strong antiproliferation in EC109 and KYSE-150 cells in a concentration-depend- ent manner.2, 4 and 8 μmol·L^-1 Jar-TTA treat- ments of EC109 cells for24 h resulted in a significant increase of early apoptosis population up to(27.9±6.1)%,(71.1±9.3)% and (65.0±9.5)%, respectively, compared to control treated cells (5.6±3.2)%.The mitochondrial OXPHOS and glycolysis were significantly inhibited in EC109 incubated by 4 and 8 μmol·L^-1 Jar-TTA for2 h.In addition, Jar-TTA induced the drop of MMP.Furthermore, the glucose uptake and the expression of GLUT4 and LDHA were distinctly inhibited in EC109 treated by Jar-TTA.Conclusions Jar-TTA induces apoptosis of human esophageal cancer cells through dual inhibition of glycolysis and oxidative phosphorylation, which is related with the drop of MMP collapse, the decrease of glucose uptake and the down-regulation of GLUT4 and LDHA in EC109 treated by Jar-TTA.
作者
范霞霞
苏楠
黄晓婧
李亚飞
贾奥
樊济配
王爱凤
赵宁民
马永成
FAN Xia-xia;SU Nan;HUANG Xiao-jing;LI Ya-fei;JIA Ao;FAN Ji-pei;WANG Ai-feng;ZHAO Ning-min;MA Yong-cheng(Clinical Pharmacology Lab, People′s Hospital, ZhengzhouUniversity, Zhengzhou 450003,China;Pharmacy Dept, Fuwai Centeral China Cardiovascular Hospital, Zhengzhou 451464, China;Collegeof Food and Biological Engineering, Henan University of Animal Husbandry and Economy, Zhengzhou 450011, China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2019年第7期950-958,共9页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81502952)
郑州大学人民医院“23456人才工程”基金
河南省医学科技攻关计划项目(No 2018020462)
关键词
二萜衍生物
食管癌
凋亡
能量代谢
糖酵解
氧化磷酸化
diterpenoid derivatives
human esophageal cancer
apoptosis
energy metabolism
glycolysis
oxidative phosphorylation
