Thiostrepton介导FOXM1下调对非小细胞肺癌细胞生长及其AG1478药物敏感性的影响  

Role of inhibition of transcription factor FOXM1 expression by Thiostrepton in the proliferation and AG1478 sensitivity of non-small cell lung cancer cell

作　　者：公小迪[1] 袁海花[1] 王炯轶[1] 郭跃辉[1] 时婧[1] 王竞[1] 姜斌[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院肿瘤科,上海201900

出　　处：《现代肿瘤医学》2013年第1期38-42,共5页Journal of Modern Oncology

基　　金：上海市教委基金资助项目(编号:08YZ47);上海市科委基金资助项目(编号:10JC1409200);上海市交通大学医学院科技基金项目(编号:11XJ22014)

摘　　要：目的:探讨硫链丝菌素(thiostrepton,TST)对肺癌细胞A549增殖及其对AG1478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检测不同浓度TST对A549细胞中FOXM1的表达影响;利用RNAi技术沉默A549细胞中FOXM1基因,RT-PCR及Westernblot检测转染效率;集落形成实验检测转染前后细胞集落形成能力的改变。结果:TST明显抑制A549细胞增殖并提高其对AG1478的药物敏感性。TST呈剂量依赖性抑制FOXM1、c-myc、CyclinB1及Bcl-2表达;与对照组相比,FOXM1 siRNA转染组的FOXM1 mRNA及蛋白表达均下降,其下游c-myc、Bcl-2的表达下降,Cleaved-PARP的表达明显上调;转染组集落数为(32.0±3.0)个,明显低于空白组及阴性对照组(NC)[集落数分别为(146.0±4.6)个、(128.7±5.7)个],P<0.05。转染组及NC组细胞集落形成抑制率分别为78.08%、11.87%,转染后细胞的集落形成能力明显降低(P<0.05);AG1478单药组的IC50值是TST与AG1478联用组的5.96倍,P<0.05。结论:TST可能通过下调FOXM1的表达抑制A549细胞的增殖、诱导其凋亡,并增加其对AG1478的药物敏感性。Objective： To investigate the change of proliferation and AG1478 sensitivity of non - small cell cancer （NSCLC） cell lines A549 with FOXM1 expression downregulated by Thiostrepton （TST）. Methods：Human NSCLC cell A549 was treated with TST or AG1478. CCK - 8 cell viability asay was performed to determine the relative growth suppression rate of A549 cell. The expression level of target genes of TST were studied by Western blot. A siRNA tar- geting FOXM1 was designed and transfected into A549 cell, an unrelated siRNA was used as negative control（NC）. RT- PCR and Western blot were used to examine the FOXM1 expression in mRNA and protein level respectively. Colony assay was employed to evaluate the colony formation ability of cell transfected by FOXM1 siRNA. Results：TST suppressed the cell proliferation and could increase chemosensitivity of A549 cell to AG1478. The expression level of FOXM1 ,c- mye ,CyclinB1 and Bcl- 2 were inhibited by TST dose -dependently. FOXM1 siRNA depleted the mR- NA and protein expressions of FOXM1 and also the expression of target genes c - myc, Bcl -2 ,while induced the ex- pression of Cleaved - PARP in A549 cell. The colony formation numbers （32.0 ± 3.0 ） were significantly fewer than those of NC and control group （ colony formation numbers were 146.0 ± 4.6,128.7 ± 5.7 respectively, P 〈 0. 05 ）. The colony formation inhibition rate of FOXM1 siRNA （78.08%）was significantly lower than that of the NC group （ 11.87 %. P 〈 0.05 ）. Conclusion： TST could significantly inhibit the proliferation and induce cell apoptosis by down -regulating the expression of FOXM1, and then increase chemosensitivity of A549 cell to AG1478.

关 键 词：非小细胞肺癌 FOXM1 细胞增殖 RNA干扰 硫链丝菌素 药物敏感性 

分 类 号：R734.2[医药卫生—肿瘤]