摘要
目的:观察结合于Bcl-xl mRNA上低结合自由能区域的反义寡核苷酸对Bcl-xl基因的抑制效应。方法:利用软件分析Bcl-xl mRNA上的低结合自由能区域,并针对这一区域合成反义寡核苷酸As587;转染高表达Bcl-xl的人乳腺癌细胞系MCF-7;实验分空白对照组、反义组(As587组)和错义组(Sc587组),转染后检测各组细胞的增殖、Bcl-xl基因及蛋白表达有无变化。结果:As587组与Sc587组相比,转染72h后细胞抑制率差异有统计学意义,P<0.01;凝胶电泳分析Bcl-xl/β-actin比值分别为0.39和0.79,P<0.05;Bcl-xl蛋白水平亦明显下降,P<0.01。结论:计算机软件辅助设计反义寡核苷酸是一种可靠的方法,以此方法设计的反义核酸As587在体外能抑制Bcl-xl基因的表达。
OBJECTIVE： To investigate the effects of an anti- sense oligodexynucleotide which targets to low free energy region of Bcl xl mRNA on anti-expression of Bcl-xl. METHODS： The low free energy region of Bcl xl mRNA was analysed by software and an ASODN, which targeted to the low free energy region of Bel-xl mRNA and named As587, was synthesized. As587was transfacted into breast cell line MCF-7 expressing Bcl-xl protein in high level. Three groups were established： blank, antisense （As587） and scramble （Sc587）. After 72 h of the transfection, the cell proliferation, Bcl-xl gene and protein expressions were observed respec tively. RESULTS： In the As587 and Sc587 groups, The cell inhibi- tion rates were significantly different （P~ 0.01 ）, Bcl-xl/^-actin ratios were 0.39 and 0. 79 （P〈0.05）, and the Bcl-xl protein decreased in the As587 group （P〈0.01）. CONCLUSION： It＇s a reliable method to use computer aided design during the application of antisense oligonucleotides, and As587 can suppress the expression of Bcl-xl in vitro. Chin J Cancer Prey Treat, 2008,15 （22） ： 1714 -- 1716
出处
《中华肿瘤防治杂志》
CAS
2008年第22期1714-1716,共3页
Chinese Journal of Cancer Prevention and Treatment
