[表达靶向生存素基因的小干扰RNA的慢病毒载体对体外培养的Hep-2细胞及裸鼠体内Hep-2细胞生长的抑制作用]
[Inhibitory effect of lentiviral vectors expressing small interfering RNA targeting survivin gene on Hep-2 cell growth in vitro and in nude mice].
作者信息
Yao Xiao-bao, Wang Xiao-xia, Zhang Shao-qiang, Yan Li-ying, Zhu Hong-liang
机构信息
Department of Otolaryngology, First Affiliated Hospital of Xi'an Jiaotong University College of Medicine, Xi'an 710061, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2010 Jan;30(1):170-2.
OBJECTIVE
To observe the effect of the lentiviral vectors expressing small interfering RNA (siRNA) for survivin gene knockdown in inhibiting Hep-2 cell growth in vitro and its tumorigenicity in nude mice.
METHODS
The tumorigenicity of Hep-2 cells transfected with the siRNA mediated by the lentiviral vectors was tested in nude mice. The expression of survivin gene of the transfected cells at the mRNA and protein levels were detected by RT-PCR and Western blotting, respectively, and the cell cycle changes were analyzed by flow cytometry.
RESULTS
Transfection of the siRNA targeting survivin significantly decreased the expression of survivin mRNA and protein in Hep-2 cells in vitro by 60%-85% and 70%, respectively, resulting also in increased cell apoptosis as shown by flow cytometry (P<0.01). The transfection significantly lowered the tumorigenicity of the cells in nude mice.
CONCLUSION
The lentiviral vectors expressing survivin siRNA can significantly inhibit survivin gene expression in Hep-2 cells and induce the cell apoptosis in vitro, and suppress the tumorigenicity of the cells in nude mice.
目的
观察表达小干扰RNA（siRNA）的慢病毒载体对生存素基因敲低在体外抑制Hep-2细胞生长及其在裸鼠体内致瘤性的影响。
方法
在裸鼠中检测经慢病毒载体介导的siRNA转染的Hep-2细胞的致瘤性。分别通过逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测转染细胞中生存素基因在mRNA和蛋白质水平的表达，并通过流式细胞术分析细胞周期变化。
结果
靶向生存素的siRNA转染显著降低了体外Hep-2细胞中生存素mRNA和蛋白质的表达，分别降低了60%-85%和70%，流式细胞术显示细胞凋亡也增加（P<0.01）。转染显著降低了细胞在裸鼠中的致瘤性。
结论
表达生存素siRNA的慢病毒载体可显著抑制Hep-2细胞中生存素基因的表达，在体外诱导细胞凋亡，并抑制细胞在裸鼠中的致瘤性。
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