鱼藤素诱导肺癌细胞凋亡和细胞周期阻滞的分子机制  

Molecular mechanism of deguelin-induced apoptosis and cell cycle arrest in lung cancer cells in vitro

作　　者：陈彦洁 柳传毅 吕满霞 方琼彤 吴新荣 CHEN Yan-jie;LIU Chuan-yi;LYU Man-xia;FANG Qiong-tong;WU Xin-rong(Dept of Pharmacy,General Hospital of Southern Theatre Command of PLA,Guangzhou 510010,China;School of Food Science and Engineering,South China University of Technology,Guangzhou 510641,China)

机构地区：[1]中国人民解放军南部战区总医院药剂科,广东广州510010 [2]华南理工大学食品科学与工程学院,广东广州510641

出　　处：《中国药理学通报》2019年第8期1109-1114,共6页Chinese Pharmacological Bulletin

基　　金：全军后勤科研计划项目(No CGZ14C003)

摘　　要：目的 探究鱼藤素是否能够阻滞细胞周期和细胞迁移,抑制非小细胞肺癌细胞的增殖。方法 鱼藤素(1.5625、3.125、6.25、12.5、25、50 μmol·L ^-1)处理H1299细胞不同时间后,CCK-8法检测细胞存活率,划痕实验测定划痕宽度及愈合率;鱼藤素(1.5、3、6 μmol·L ^-1)处理24 h,PI单染测定细胞周期,Annexin V-FITC/PI双染检测凋亡率,qPCR检测药物处理后的H1299细胞中CDK4、CDK6和CCND1的基因表达情况。结果 鱼藤素能够降低H1299细胞的存活率,作用24、48、72 h的IC 50 值分别为(5.47±0.97)、(4.01±0.45)、(2.86±0.19)μmol·L ^-1 ,并抑制细胞的迁移与愈合能力。PI单染流式细胞术检测到其能阻滞细胞周期,使细胞阻滞在G 0/G 1期,流式双染实验测得其可诱导细胞的凋亡,qPCR发现鱼藤素可下调CDK4、CDK6和CCND1的基因表达。结论 鱼藤素可抑制H1299细胞的增殖,抑制细胞的运动迁移能力,阻滞细胞周期,诱导细胞凋亡,且鱼藤素可通过下调细胞周期调控系统中的CDK4、CDK6、CCND1基因表达来调控细胞周期,达到抗肿瘤作用。Aim To explore whether deguelin can block cell cycle and cell migration,inhibit the proliferation of non-small cell lung cancer cells.Methods H1299 cells were treated with 1.562 5,3.125,6.25,12.5,25,50 μmol·L^-1 deguelin for different time (24,48,72 h);cell viability was detected by CCK-8 assay,and cell migration ability was tested by scratch assay.H1299 cells were treated with 1.5,3,6 μmol·L^-1 deguelin for 24 h.Flow cytometry with PI single staining and Annexin V-FITC/PI double staining experiment were used to evaluate cell cycle and apoptosis.qPCR was used to detect the regulatory effects of deguelin and its carbamate derivative on the Cyclin D-CDK4/6 complex at the gene level.Results Deguelin inhibited cell growth and the IC 50 value of deguelin was (5.47±0.97),(4.01±0.45),(2.86±0.19)μmol·L^-1 when treated with 24,48,72 h respectively.Deguelin also inhibited the healing ability of H1299 cells and the migration of H1299 cells significantly (P<0.05).Deguelin could block H1299 cell cycle in G 1 phase.Flow cytometry combined with Annexin V-FITC/PI double staining showed that deguelin could induce apoptosis of H1299 cells.qPCR experiments showed that deguelin could down-regulate the expression of CDK4,CDK6 and Cyclin D1 genes significantly (P<0.05).Conclusions Deguelin may regulate cell cycle by down-regulating CDK4,CDK6 and Cyclin D1 genes in the cell cycle regulation system,and reduce the migration ability of tumor cells to induce apoptosis.

关 键 词：鱼藤素 H1299细胞 细胞周期 抗肿瘤 CYCLIN D1 CDK4/CDK6 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]