天然二萜衍生物JYD01促进己糖激酶Ⅱ与线粒体解离诱导胃癌细胞凋亡  

JYD01,a newly synthesized diterpenoid derivative,induces apoptosis in human gastric cancer cells through detaching hexokinaseⅡfrom mitochondria

作　　者：苏楠[1] 范霞霞 王爱凤[2] 马永成[2] SU Nan;FAN Xia-xia;WANG Ai-feng;MA Yong-cheng(College of Food and Biological Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450011,China;Dept of Pharmacy,Henan Provincial People’s Hospital,Dept of Pharmacy of Centeral China Fuwai Hospital,Centeral China Fuwai Hospital of Zhengzhou University,Zhengzhou 450003,China)

机构地区：[1]河南牧业经济学院食品与生物工程学院,河南郑州450011 [2]河南省人民医院药学部,华中阜外医院药学部临床药理室,郑州大学华中阜外医院,河南郑州450003

出　　处：《中国药理学通报》2021年第6期871-877,共7页Chinese Pharmacological Bulletin

基　　金：中华医学会临床药学分会吴阶平医学基金会科研项目(No 320.6750.19090-4);国家自然科学基金资助项目(No 81502952);河南省高等学校重点科研项目(No 21A330002);河南省重点研发与推广专项(No 192102310155);河南省医学科技攻关计划项目(No 2018020462)。

摘　　要：目的研究二萜衍生物JYD01诱导己糖激酶Ⅱ(hexokinaseⅡ,HKⅡ)与线粒体解离作用,并探讨其抗胃癌细胞增殖机制。方法MTT法检测JYD01对胃癌细胞MGC-803、BGC-823的抑制作用;细胞能量代谢分析仪实时检测细胞糖酵解;流式细胞术定量分析细胞凋亡及线粒体膜电位(mitochondrial membrane potential,MMP)、荧光显微镜进行细胞形态学研究及MMP观察;Western blot检测蛋白表达。结果JYD01明显抑制MGC-803、BGC-823细胞增殖,且呈浓度依赖性;JYD01可以促使HKⅡ与线粒体解离,抑制胃癌细胞糖酵解,诱导MMP下降并导致细胞色素C由线粒体释放至细胞质;1、2、4μmol·L-1 JYD01诱导MGC-803总凋亡率分别为(32.2±4.7)%、(46.5±6.3)%和(66.4±7.5)%,与对照组(5.8±2.3)相比,差异均具有显著性(P<0.01)。结论JYD01明显抑制胃癌细胞生长,机制可能与其促进HK2与线粒体解离,从而抑制糖酵解功能,并启动线粒体凋亡途径有关。本研究为天然二萜化合物的抗肿瘤研究提供了新的视角。Aim To investigate the effect of JYD01,an ent-kaurane diterpenoid analog,on detaching hexokinaseⅡ(HKⅡ)from mitochondria,and discuss the underlying mechanism of anti-gastric cancer cell proliferation.Methods MTT assay was performed to measure the effect of JYD01 on the growth capacity of human gastric cancer cell lines MGC-803 and BGC-823.The glycolysis of MGC-803 cells in response to JYD01 was analyzed using a Seahorse XFp extracellular flux analyzer by real-time measurements of the extracellular acidification rate(ECAR,indicative of glycolysis).The effect of JYD01 in mitochondrial membrane potential(MMP)and apoptosis was observed by a fluorescence microscopy.The apoptotic rate and the quantitative analysis of MMP falling of cell lines treated with JYD01 were analyzed by flow cytometry.The pro teins were determined by Western blot.Results JYD01 observably inhibited the growth of MGC-803 and BGC-823 cells in a dose-dependent manner.JYD01 induced a dose-dependent detachment of HKⅡfrom mitochondria of MGC-803 cells,effectively reduced glycolysis,and caused the drop of MMP leading to the release of cytochrome c.1,2 and 4μmol·L-1 JYD01 treatments of MGC-803 cells for 24 h,resulted in a significant increase of total apoptosis population up to(32.2±4.7)%,(46.5±6.3)%and(66.4±7.5)%,respectively,compared to control treated cells(5.8±2.3)%,and the differences were all significant.Conclusions The new compound JYD01 has a strong anti-gastric cancer activity,which is related with the detachment of HKⅡfrom mitochondria,glycolysis inhibition,and the activation of mitochondria apoptosis pathway in MGC-803 cells treated by JYD01.This study provides a new perspective for the anti-tumor research of natural ent-kaurene diterpenoids.

关 键 词：天然二萜衍生物 HKⅡ 糖酵解 线粒体 抗肿瘤 凋亡 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]