靶向survivin基因小干扰RNA设计及鉴定

SiRNA designing and identification for silencing survivin protein expression

目的:利用生物信息学技术预测设计特异抑制survivin蛋白表达的siRNA序列,瞬时转染食道癌细胞KYSE-70,并观察对survivin蛋白表达的沉默效果。方法:依据NCBI数据库获得的survivin mRNA序列,利用Whitehouse,Sidirect和Rational SiRNA Design软件预测设计siRNA干扰序列,并利用BLAST分析干扰序列与人类全基因组的同源性;转染食管癌细胞KYSE-70,Western blot法鉴定蛋白表达。结果:初步预测出3条含21个核苷酸的survivin干扰序列,瞬时转染食道癌细胞KYSE-70后survivin蛋白表达明显下降。结论:运用生物信息学技术可快速便捷预测出抑制survivin表达的siRNA序列,为研究survivin在食管癌发生发展中的作用奠定了基础。

Objective： To predict and design the siRNA sequence by using the bio-informatics technology for inhibition of survivin protein expression, observe the effects of transfecting siRNA into e- sophageal cancer cell line KYSE-70 on silencing survivin protein expression. Methods： According to the survivin sequence obtained from NCBI database, Whitehouse, Sidirect and Rational SiRNA Design software were used to predict the siRNA interference sequence and followed by homological analysis with BLAST. SiRNA was transfected into esophageal cancer cell line KYSE-70. Survivin protein expression was examined by western blot. Results： 3 survivin interfere RNA sequences in- cluding 21 nucleotides were predicted. Western blot showed that the expression of survivin protein in KYSE-70 cells decreased significantly after transfection with siRNA. Conclusion ： The bio-infor- matics technology can be conveniently employed for survivin siRNA design which could promote the further study on the occurrence and development of esophageal cancer.