Survivin反义寡核苷酸诱导肺癌细胞株凋亡的研究  

The synergic effect of survivin antisense oligonucleotide on apoptosis in lung cancer cell lines

作　　者：夏雪梅[1] 陈余清[1] 蔡映云[2] 李殿明[1] 胡俊锋[1] 黄礼年[1] 

机构地区：[1]蚌埠医学院附属医院呼吸病科,安徽蚌埠233004 [2]复旦大学附属中山医院肺科,上海200032

出　　处：《蚌埠医学院学报》2005年第1期4-8,共5页Journal of Bengbu Medical College

基　　金：安徽省自然科学基金资助项目 (0 3 0 43 70 5 )

摘　　要：目的 :探讨生存素 (Survivin)的反义寡核苷酸 (ASODN)对肺癌细胞株的作用。方法 :在脂质体介导下 ,分别以10 0nmol/L、30 0nmol/L和 5 0 0nmol/L的SurvivinASODN作用于小细胞肺癌细胞株NCI H4 4 6和肺腺癌细胞株SPC A1,于2 4h、4 8h、72h用逆转录聚合酶链反应 (RT PCR)检测SurvivinmRNA表达 ,Westernblot检测Survivin蛋白 ,流式细胞仪检测凋亡。结果 :NCI H4 4 6和SPC A1皆表达Survivin基因和蛋白。随着ASODN浓度增加 ,SurvivinmRNA明显下调 ,其差异有显著性 ,其中ASODN 5 0 0nmol/L作用 72h时两种细胞SurvivinmRNA抑制率分别达 6 2 .72 %和 6 7.4 3% ;Survivin蛋白表达也有类似的变化 ;ASODN对两种细胞生长抑制和诱导凋亡作用随浓度增大和作用时间延长而增强。结论 :SurvivinASODN能够抑制肺癌细胞株Survivin基因和蛋白表达 ,呈时间、浓度依赖性 ,诱导凋亡的效果也呈剂量、时间依赖性 ,5 0 0nmol/L的浓度作用 72h效果最佳。Objective:To investigate effect of Survivin antisense oligonucleotide(ASODN) on expression of Survivin mRNA and qrotein,and apoptosis in lung cancer cell lines.Methods:The lung cancer cell lines,NCI-H446 and SPC-A1 were transfected with Survivin ASODN (100 nmol/L,300 nmol/L,500 nmol/L),NSODN(100 nmol/L,300 nmol/L,500 nmol/L),by using Lipofectin for 24 hour,48 hour and 72 hour.The expression of Survivin mRNA was measured by RT-PCR technique.The expression of Survivin protein was measured by western blotting assay,the apoptosis index,proliferation index and cell cycle were measured by flow cytometry (FCM).Results:Survivin ASODN induced downregulation of the Survivin mRNA of NCI-H446 and SPC-A1 cells in dose-dependent and time-dependent manners,and at concentration of 500 nmol/L for 72 hour,the suppression rate of Survivin mRNA was 62.72% and 67.43%,respectively.Survivin ASODN also induced downregulation of the Survivin protein in both lung cancer cell lines.By analyzing the cell cycle and apoptosis,it was demonstrated that G 0/G 1 and AI were increased and apoptosis was induced with dose-dependent and time-dependent manners.Its maximum effect was achieved at a concentration of 500 nmol/L for 72 hour,at which apoptosis indexes were 44.71% and 31.83%,respectively.Conclusions:Both NCI-H446 and SPC-A1 express Survivin mRNA and protein;Survivin ASODN can downregulate Survivin mRNA and protein;Survivin ASODN can induce apoptosis in dose-dependent and time-dependent manners,its maxinum effect is achieved at a concentration of 500 nmol/L for 72 hour.

关 键 词：肺肿瘤 生存素 反义寡核苷酸 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]