STAT3-siRNA表达框架的设计与构建

Design and construction of STAT3-siRNA

【目的】设计和构建针对STAT 3基因的siRNA表达框架,并在结直肠癌细胞中观察其干扰效果。【方法】GeneBank中查询STAT 3基因序列,Oligo 6.0软件设计siRNA靶序列,合成并修饰靶序列后PCR扩增siRNA表达框架,通过脂质体介导,直接将扩增产物转染至结直肠癌细胞SW480中,48 h后提取细胞总RNA进行RT-PCR检测干扰效果。【结果】RT-PCR结果显示SW480细胞内靶基因STAT 3 mRNA水平下降(62.16±12.50)%。【结论】本实验成功设计并构建的siRNA表达框架能干扰人结直肠癌SW480细胞内STAT 3基因的表达。

[Objective] To construct a siRNA expression cassette targeted to gene STAT3 and observe the efficiency to RNA interference in colorectal cancer cell. [Methods] The gene order of STAT3 was inquired in GeneBank and design the target sequence of siRNA aimed at STAT3 gene using software Oligo 6.0. After synthesizing and modifying the target sequence, the siRNA expression cassette was amplified by polymerase chain reaction （PCR）. Mediated by liposome, the amplified product was transfected into colorectal cancer cell SW480. After 48 hours, the total RNA was extracted for reverse transcriptional-polymerase chain reaction （RT-PCR） to detect the efficiency of RNA interference. [Results] The STAT3 gene expression was decreased （62.16 ±12.50）% at the mRNA level in comparison with control cells demonstrated by RT-PCR detection. [Conclusions] We successfully construct a siRNA expression cassette targeted to gene STAT3 and it can interference the expression of STAT3 gene in colorectal cancer SW480.