雷公藤甲素丁二酸单酯对A549/DDP细胞的抑制作用研究  

Antitumor Activities of Triptolide Succinate Monoester on A549/DDP Cells

作　　者：杨俊 王丹丹 梁玉清 丁丽娜[1] 卓俊睿 梁光平 YANG Jun;WANG Dan-dan;LIANG Yu-qing;DING Li-na;ZHUO Jun-rui;LIANG Guang-ping(Zunyi Medical and Pharmaceutical College,Zunyi 563006,China;Tongren Polytechnic College,Tongren 554300,China)

机构地区：[1]遵义医药高等专科学校,贵州遵义563006 [2]铜仁职业技术学院,贵州铜仁554300

出　　处：《化学试剂》2022年第8期1128-1135,共8页Chemical Reagents

基　　金：遵义市科技计划课题项目(遵市科合社字[2018]46号);贵州省科技计划项目(黔科合基础[2020]1Y397号)。

摘　　要：研究雷公藤甲素丁二酸单酯(YJ-4)对A549/DDP抑制作用及其机制。以雷公藤甲素为原料,与丁二酸酐在DMAP/Pyridine催化下反应后得到化合物YJ-4,其结构经^(1)HNMR,^(13)CNMR和HR-ESI-MS表征,采用MTT法评价其对人非小细胞肺癌细胞(A549)、人肺腺癌耐顺铂株(A549/DDP)、人乳腺癌细胞(MCF-7)的体外抗肿瘤活性;不同浓度的化合物YJ-4作用于A549/DDP细胞后,DAPI染色法检测细胞核状态,PI染色法检测细胞周期,Annexin V-FITC/PI双染法检测细胞凋亡;分子对接软件AutoDock来评价化合物YJ-4与MDM2蛋白结合的能力。化合物YJ-4对A549/DDP((33.32±3.55)μmol/L)、A549((4.31±1.01)μmol/L)、MCF-7((16.08±2.63)μmol/L)细胞株均具有良好的细胞毒性,是阳性对照药顺铂的数倍,且通过阻滞A549/DDP细胞周期G_(2)/M期,从而诱导细胞凋亡,与MDM2的GLY-16氨基酸残基形成氢键(d=1.9),结合能(affinity)为-8.4 kcal/mol,能够较好的结合。化合物YJ-4对耐药细胞株A549/DDP细胞具有良好的细胞毒活性,其作用机制为促进A549/DDP细胞凋亡、阻滞细胞周期G_(2)/M期,可能与抑制MDM2蛋白和p53相互作用有关。To investigate the effects and mechanisms of triptolide succinate monoester(YJ-4)on cisplatin-resistant human lung adenocarcinoma cells(A549/DDP).Compound YJ-4 was synthesized by catalyzing triptolide and succinic anhydride under DMAP/pyridine.Its structure was characterized by ^(1)HNMR,^(13)CNMR,and HR-ESI-MS.Its anti-tumor activity in vitro against human non-small cell lung cancer cells(A549),human breast cancer cells(MCF-7),and A549/DDP cells were evaluated using the MTT method.After A549/DDP cells were treated with YJ-4 at different concentrations,the nuclear state was detected by DAPI staining,the cell cycle was detected by PI staining,and the apoptosis was detected by Annexin V-FITC/PI double staining.The binding ability of YJ-4 to MDM2 protein was evaluated using molecular docking software AutoDock.Compound YJ-4 presented high cytotoxicity to A549/DDP((33.32±3.55)μmol/L),A549((4.31±1.01)μmol/L),and MCF-7((16.08±2.63)μmol/L)cells,which was significantly superior to the positive control drug cisplatin.Additionally,YJ-4 blocked the G_(2)/M phase of the A549/DDP cell cycle and induced apoptosis.It could form a hydrogen bond with amino acid residue GLY-16 of MDM2(d=1.9),with an affinity of-8.4 kcal/mol,suggesting a high binding ability.Compound YJ-4 had a high cytotoxic activity to drug-resistant cell line A549/DDP.Its mechanism was to promote A549/DDP cell apoptosis and block the G_(2)/M phase of the cell cycle,which may be related to the inhibition of the interaction between MDM2 protein and p53.

关 键 词：雷公藤甲素丁二酸单酯 衍生物 人肺癌细胞耐顺铂株A549/DDP 体外抗肿瘤活性 

分 类 号：R914.4[医药卫生—药物化学]