摘要
目的:利用基因转染和RNA干涉(RNAi)技术,在SPC-A1细胞中建立Livin异构体α和β特异的基因沉默体系,探讨其在诱导SPC-A1细胞凋亡效应中的不同作用及价值。方法:在肺腺癌SPC-A1细胞中稳定表达Livinα+β、Livinα和Livinβ特异性小干涉RNA(siRNA),观察SPC-A1细胞形态、增殖等生物学行为的改变,TUNEL法和流式细胞仪检测细胞凋亡率。结果:Livinα+β、Livinα和Livinβ基因沉默后,SPC-A1细胞克隆形成能力较对照组显著下降(P<0.05);转染后SPC-A1细胞发生凋亡,TUNEL法显示细胞凋亡指数分别为(9.20±1.90)%、(6.75±1.20)%和(4.82±1.20)%,(流式细胞仪检测)细胞凋亡率分别为(7.48±1.30)%、(5.50±0.90)%和(4.16±0.70)%。结论:Livinα和β两种异构体均能作为诱导肺癌细胞凋亡的分子靶点,Livin基因沉默靶向诱导肺癌细胞凋亡可能作为手术、化疗、放疗等传统治疗的辅助手段。
Purpose： To explore different functions and values of Livin α and Livin β-specific silencing in inducing SPC-A1 cell apoptosis, respectively. Methods： First, Livin α ＋ β, Livinαand Livin β-specific siRNA were steadily expressed in SPC-A1 respectively. Next, after isoform-spccific gene silencing, biological changes with regard to cell morphology and proliferation of lung cancer cell were observed. Then, TUNEL and FCAS were performed to test the rate of apoptosis. Lastly, statistical analysis was performed to explore the different functions and values of isoform-specific gene silencing in inducing lung cancer cell apoptosis. Results： After gene silencing of Livin α＋ β, Livin α and Livin β, there was an evident decrease in colony forming ability of SPC-A1 compared with the control（ P 〈 0.05）. Transfected SPC-A1 had apoptosis. TUNEL showed apoptosis index as （9.20±1.90） %, （6.75 ±1.20） % and （4.82 ± 1.20） %, respectively. FCAS shuwed rate of apoptosis as （7.48 ± 1.30） %, （5.50 ± 0.90） % and （ 4.16 ± 0.70） %, respectively. Conclusions： Livin α and β can be used as targets in inducing lung adenocarcinoma cells apoptosls. Livin gene silencing is a hopeful new method for lung cancer treatment, serving as an adjuvant therapy for operation, chemotherapy and radiotherapy of lung cancer.
出处
《中国癌症杂志》
CAS
CSCD
2005年第6期505-509,共5页
China Oncology
基金
国家自然科学基金资助青年项目(No:30200282)。
关键词
凋亡抑制蛋白
LIVIN
凋亡
基因沉默
RNA干涉
非小细胞肺癌
inhibitor of apoptosis family of protein
Livin
apoptosis
gene-silencing
RNA interference,non-small-cell lung cancer
