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

基础医学与临床 ›› 2007, Vol. 27 ›› Issue (12): 1365-1369.

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

刘黎明陈昌杰陈正徐段光军胡华成

苏州大学附属第二医院呼吸内科蚌埠医学院检验系蚌埠医学院检验系苏州大学附属第二医院呼吸内科苏州大学附属第二医院呼吸内科

收稿日期:2007-02-26 修回日期:2007-06-04 出版日期:2007-12-25 发布日期:2007-12-25
通讯作者: 胡华成

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

Li-ming LIU, Chang-jie CHEN, Zheng-xu CHEN, Guang-jun DUAN, Hua-cheng HU

Dept of Respiratory Medicine, the Second Affiliated Hospital, Soochow University

Received:2007-02-26 Revised:2007-06-04 Online:2007-12-25 Published:2007-12-25
Contact: Hua-cheng HU

摘要/Abstract

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、免疫印迹法（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%，免疫印迹法显示蛋白表达下降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。

Abstract: 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 (MTT)colorimetry and flow cyclometry. Results SURVIVIN -siRNA expression vector was constructed and transfected into A549 cell successfully. It couldeffectively reduce the mRNA and protein level of SURVIVIN (76.4% and 84.5% respectively).

刘黎明陈昌杰陈正徐段光军胡华成. 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[J]. 基础医学与临床, 2007, 27(12): 1365-1369.

Li-ming LIU; Chang-jie CHEN; Zheng-xu CHEN; Guang-jun DUAN; Hua-cheng HU. Construction of SURVIVIN specific small interfering RNA expression vector and its antiproliferative and proapoptotic effect on lung adenocarcinoma[J]. Basic & Clinical Medicine, 2007, 27(12): 1365-1369.