靶向BIRC5,Bcl-2siRNA慢病毒表达载体最佳干扰序列的筛选

Effective Jamming Sequence Screening of the Expression Vector in Slow Virus of the Target to the siRNA of BIRC5 and Bcl-2

目的 针对设计合成构建的BIRC5和Bcl-2小分子干扰RNA（siRNA）的慢病毒质粒表达载体进行有效靶序列筛选,探讨其对靶基因的封闭和抑制作用.方法 将BIRC5和Bcl-2 siRNA慢病毒质粒表达载体通过脂质体介导,分别转染体外培养的Tca-8113细胞.采用半定量RT-PCR检测转染细胞BIRC5和Bcl-2 mRNA表达及Western blot检测蛋白表达水平.结果 BIRC5和Bcl-2 mRNA抑制率分别为35%～76%和30%～72%,所设计的siRNA序列不同程度降低和下调其mRNA及相应的蛋白表达,实验组与对照组比较差异有统计学显著性意义（P〈0.05）,阴性对照组无此作用（P〉0.05）.结论 设计合成的siRNA干扰序列能抑制其靶基因mRNA和蛋白的表达,不同的干扰靶点其抑制效果不同,本实验成功筛选出两对最佳干扰序列作为对肿瘤细胞基因治疗的实验研究.

Objective Through screening effect target sequence in the slow viral material particle expression vector which carry on the design and synthesis of the BIRC5 and Bcl-2 small molecules disturbs RNA （siRNA）,object was to research its seal and inhibitory action in the target gene. Methods The BIRC5 and Bcl-2 siRNA in the slow virus material particle expression vector was transferred into the vitro person lingual cancer cells Tca-8113 through the lead of llpin body. The examination to the expression of BIRC5 and Bcl-2 mRNA in transfer cells was throngh the method of RT-PCR,and the examination to the expression of protein was Western blot. Results The inhibitory rates of BIRC5 and Bcl-2 mRNA were 35%- 76% and 30%-72% respectively. All the siRNA sequence could reduce and down regulate the expression of protein. At all the level,and there was significant in statistics with the comparison between the experimental group and the control group（ P 〈0. 05） ,however the negative control group was noneffective（P〉0. 05）. Conclusion The synthetical siRNA disturbs sequence could restrain the expression of target gene mRNA and protein,and the repressive impression varied in different target sequence. Have triumphantly obtained two effective disturbs sequence to research the therapy of lingual cancer.