AZD8055抑制胆管癌细胞HuCCT1的迁移及EMT进程的机制研究  

Study on mechanism of AZD8055 inhibiting migration and EMT progression of cholangiocarcinoma cell HuCCT1

作　　者：王雪 毕宇宁[1,3] 闫文帝 张鑫 董颖 匡子藤[1,3] 林贞花 任香善[1,2,3] WANG Xue;BI Yu-ning;YAN Wen-di;ZHANG Xin;DONG Ying;KUANG Zi-teng;LIN Zhen-hua;REN Xiang-shan(Cancer Research Center,Yanbian University,Yanji Jiling 133002,China;Dept of Pathology,Yanbian University Medical College,Yanji Jiling 133002,China;Key Lab of the Science and Technology Dept of Jilin Province,Yanji Jiling 133002,China)

机构地区：[1]延边大学肿瘤研究中心,吉林延吉133002 [2]延边大学医学院病理学教研室,吉林延吉133002 [3]延边大学吉林省科技厅重点实验室,吉林延吉133002

出　　处：《中国药理学通报》2019年第10期1429-1436,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 31460303,81660609);吉林省科技厅自然科学基金项目(No 20180101007);吉林省教育厅自然科学基金项目(No JJKH20191116KJ)

摘　　要：目的探讨哺乳动物雷帕霉素靶蛋白(mTOR)双重抑制剂AZD8055在抑制人胆管癌细胞HuCCT1的迁移及EMT进程中的作用及分子机制。方法MTT法与平板克隆形成实验检测AZD8055对胆管癌细胞增殖的影响;划痕愈合实验和Transwell小室迁移实验检测AZD8055对HuCCT1细胞迁移能力的影响;Western blot法检测EMT标志相关蛋白、Akt/mTOR信号通路蛋白及DEK蛋白的表达;利用STITCH、GeneMANIA数据库,分析AZD8055、DEK、Akt信号通路相互作用关系;在DEK基因沉默后,检测胆管癌细胞增殖活力、迁移能力及Akt/mTOR信号通路相关蛋白表达水平的变化。结果AZD8055可抑制胆管癌细胞的增殖及迁移能力,同时抑制Akt/mTOR信号通路相关蛋白、DEK蛋白表达及EMT的进程;沉默DEK基因可明显抑制胆管癌细胞增殖及迁移能力,并降低Akt、S6、4EBP1蛋白的磷酸化水平。结论AZD8055抑制HuCCT1细胞的迁移及EMT进程,其机制与下调DEK,抑制Akt/mTOR信号通路有关。Aim To investigate the molecular mechanisms of the dual inhibitor of mammalian rapamycin target protein(mTOR) AZD8055 in migration and EMT process inhibition of the human cholangiocarcinoma cell line HuCCT1.Methods The viability of HuCCT1 cells treated with different concentrations of AZD8055 was measured by MTT assay,and the colony formation ability of HuCCT1 was detected by colony formation assay.The effect of AZD8055 on the motility of HuCCT1 cells was examined by wound healing assay and Transwell assay.The expression levels of the protein associated with Akt/mTOR pathway,epithelial-mesenchymal transition(EMT) process and DEK were detected by Western blot.The interaction relationship between AZD8055,DEK and Akt signaling pathway was analyzed by STITCH and GeneMania databases.Cholangiocarcinoma cells′proliferation,migration capacities and Akt/mTOR signaling pathway-related protein expression levels were detected after DEK gene silencing.Results Compared with control group,AZD8055 inhibited the proliferation and migration capacities of cholangiocarcinoma cells,and suppressed the expression levels of Akt/mTOR signaling pathway-related markers,down-regulated DEK expression and inhibited EMT process.DEK silence significantly inhibited cell proliferation,migration and significantly decreased the phosphorylation levels of Akt,S6,and 4EBP1.Conclusions AZD8055 treatment inhibits the migration and EMT progression of HuCCT1 cells,and its mechanism is associated with DEK down-regulation and inhibition of Akt/mTOR signaling pathway.

关 键 词：AZD8055 胆管癌 DEK 上皮-间质转化 增殖 迁移 Akt/mTOR信号通路 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]