转染siRNA特异性下调人主动脉平滑肌细胞PAI表达

Transfect PAI siRNA into human aorta smooth muscle cells

目的转染化学合成siRNA特异性下调人主动脉平滑肌细胞PAI表达。方法人工合成一对互补并编码相应短发夹状PAI小核糖核酸,体外退火;采用电转染方法将小RNA导入人主动脉平滑肌细胞中,采用逆转录聚合酶链反应(RT-PCR)、western blot法分别检测转染细胞PAI mRNA和蛋白的表达变化。结果转染PAI siRNA载体后,人主动脉平滑肌细胞PAI mRNA和蛋白表达均较对照组明显下降(P<0.01)。结论转染人主动脉平滑肌细胞后,并可快速特异性地封闭PAI的表达,为进一步研究PAI作用机制提供了实验基础。

Objective To construct short interfering RNA （siRNA） eukaryotic expression vector for PAI and to transfect into human aorta smooth muscle cells（HASMC） .Methods A PAI siRNA targeting human mRNA common sequence was synthesized. The siRNA was transfected into HASMC by electROporation. Results The expressions of PAI mRNA and protein in HASMC cells of experiMental groups were significantly decreased, compared to that of controls （ P 〈 0.01 ）. Conclusion It indicated that hairpin siRNA eukaryotic expression vector for PAI would be successfully estabhshed, and it might play a specific inhibitory role in the cell hnes. At the same time we obtained ceils with the stable expression of PAI siRNA. This study laid experimental foundation for further research of the therapy of PAl siRNA vector.