国际肿瘤学杂志

国际肿瘤学杂志 ›› 2014, Vol. 41 ›› Issue (2): 135-140.doi: 10.3760/cma.j.issn.1673-422X.2014.02.017

• 论著 • 上一篇    下一篇

硫链丝菌肽下调FOXM1的表达对肺癌细胞生长及凋亡的影响

公小迪, 袁海花, 王炯轶, 郭跃辉, 孔飞飞, 姜斌   

  1. 201900 上海交通大学医学院附属第三人民医院肿瘤科
  • 收稿日期:2013-05-19 修回日期:2013-10-20 出版日期:2014-02-08 发布日期:2014-01-26
  • 通讯作者: 姜斌，E-mail:dr_jiang@yeah.net E-mail:dr_jiang@yeah.net
  • 基金资助:

    上海市教委基金（08YZ47）；上海市科委基金（10JC1409200）

Role of FOXM1 expression inbibition regulated by thiostrepton in the proliferation and apoptosis of lung cancer cells

Gong Xiaodi, Yuan Haihua, Wang Jiongyi, Guo Yuehui, Kong Feifei, Jiang Bin   

  1. Department of Oncology, Third People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 201900, China
  • Received:2013-05-19 Revised:2013-10-20 Online:2014-02-08 Published:2014-01-26
  • Contact: Jiang Bin E-mail:dr_jiang@yeah.net

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摘要： 目的 探讨硫链丝菌肽（TST）对肺癌细胞A549增殖、凋亡及其对AG1478(EGFR-TKI)药物敏感性的影响。方法 CCK8法检测TST、AG1478单药及两药联用后A549细胞的增殖抑制率；Western blot检测TST对叉头转录因子M1(FOXM1)表达的影响；Caspase3比色测定法检测TST对Caspase-3活化程度的影响；利用RNAi技术沉默A549细胞中FOXM1基因，检测FOXM1及增殖凋亡相关分子表达变化。结果 TST增加A549细胞对肺癌靶向药物AG1478的敏感性，其IC50值由(4.35±0.45) μmol/L降至(0.73±0.05) μmol/L（t=11.02，P＜0.05）；TST抑制FOXM1及下游cMyc、Cyclin B1、Bcl-2分子的表达，上调P21、Cleaved Caspase-3、Cleaved PARP表达的同时，呈时间及剂量依赖性诱导Caspase3分子活化；沉默FOXM1后其下游分子cMyc、Cyclin B1、Bcl-2、P21及Cleaved PARP的表达改变同TST作用后相似，细胞中Cleaved Caspase-3的荧光表达明显增多。结论 TST介导的FOXM1下调可抑制A549细胞增殖，诱导其凋亡，并增加其对AG1478的药物敏感性。

关键词: 肺肿瘤, RNA干扰, 叉头转录因子M1, 硫链丝菌肽, 药物敏感性

Abstract: ObjectiveTo investigate the effect of thiostrepton (TST) on cell proliferation, apoptosis and the chemosensitivity of nonsmall cell cancer (NSCLC) cell lines A549 to AG1478 (EGFRTKI). MethodsNSCLC cell A549 was treated with TST, AG1478 or the combination of the two drugs. CCK8 cell viability assay was performed to determine the relative growth suppression rate of A549 cell. The expression level of forkhead transcription factors M1 (FOXM1) was studied by western blot. Caspase3 colorimetric assay was employed to evaluate the effect of TST on Caspase3 activity. A siRNA targeting FOXM1 was designed and transfected into A549 cells. RTPCR and western blot were used to examine the expressions of FOXM1 and proliferation and apoptosis related molecules. ResultsTST increased the chemosensitivity of A59 cells to AG1478, a kind of moleculartargeted agent to advanced lung cancer, which made the IC50 value reduce from (4.35±0.45) μmol/L to (0.73±0.05) μmol/L (t=11.02, P＜0.05). Besides, TST inhibited the expressions of FOXM1 and its effectors including cMyc, Cyclin B1 and Bcl2, while upregulated P21, Cleaved Caspase3 and Cleaved PARP. Meanwhile, TST improved the activity of Caspase3, which showed time and dosedependent effects. FOXM1 siRNA changes the expression of the downstream effectors such as cMyc, Cyclin B1, Bcl2, P21 and  Cleaved PARP, which was similar to the effect of TST. Also, the fluorescent expression of Cleaved Caspase3 significantly increased in A549 cells. ConclusionTST can significantly inhibit proliferation and induce apoptosis by downregulating the expression of FOXM1, and then increase chemosensitivity of A549 cell to AG1478.

Key words: Lung neoplasms, RNA interference, Forkhead transcription factors M1, Thiostrepton, Drug sensitivity