Survivin特异小干扰RNA表达载体构建及对肺腺癌细胞增殖和凋亡的影响  

Construction of Survivin specific small interfering RNA expression vector,its antiproliferative and proapoptotic effect on lung adenocarcinoma cells

作　　者：刘黎明[1] 陈昌杰[2] 陈正徐[2] 段光军[1] 胡华成[1] 

机构地区：[1]苏州大学第二附属医院呼吸内科,江苏苏州215004 [2]蚌埠医学院检验系,安徽蚌埠233004

出　　处：《基础医学与临床》2007年第12期1365-1369,共5页Basic and Clinical Medicine

摘　　要：目的测定Survivin特异小干扰RNA(siRNA)表达载体对肺腺癌细胞A549 Survivin基因表达的抑制及对增殖和凋亡的影响。方法构建Survivin特异性小干扰RNA表达载体,转染A549细胞,筛选出稳定表达的细胞株。用荧光实时定量反转录聚合酶链反应法、Western blot和免疫组化法检测Survivin基因mRNA和蛋白的表达水平,四甲基偶氮唑蓝(MTT)法检测细胞活性,流式细胞仪测定细胞凋亡率。结果经测序鉴定证实成功构建Survivin-siRNA表达载体。Survivin基因mRNA表达量下降76.4%,Western blot显示蛋白表达下降84.5%,免疫组化法也显示Survivin蛋白表达下降。MTT法测定细胞增殖较对照组减慢(P<0.01),细胞凋亡率较对照组增加8.95%(P<0.01)。结论Survivin特异siRNA表达载体下调Survivin基因表达,抑制A549细胞的增殖,促进凋亡。Objective To test the antiproliferative and proapoptotic effect of Survivin small interfering RNA （siRNA） expression vector on lung adenocarcinoma cell A549 in vitro. Methods The Survivin-siRNA expression vector was constructed and conformed by sequencing. The inhibitory effect of Survivin-siRNA was tested by fluorescent quantitative reverse transcription polymerase chain reaction （FQ RT-PCR） , Western blot and immunohistochemistry. The cell proliferation and apoptotic rate were assayed by tetrazolium bromide （MTF） colorimetry and flow cytometry. Results Survivin -siRNA expression vector was constructed and transfected into A549 cell. It effectively reduced mRNA and protein level of Survivin. Immunohistochemistry also showed lower expression of Survivin. The A549 cells transfected with Survivin-siRNA had a lower cellular growth rate than that of control group （ P 〈 0. 01 ）,apoptotic rate increased by 8.95% than that of control group （ P 〈 0. 01 ）. Conclusion The Survivin-siRNA expression vector can inhibit the expression of Survivin gene and the proliferation of A549 cell. It can also effectively induce the apoptosis of lung adenocarcinoma cell A549.

关 键 词：小干扰RNA SURVIVIN 肿瘤细胞系 A549细胞 

分 类 号：R734.2[医药卫生—肿瘤]