CBP基因沉默抑制血管平滑肌细胞增殖的研究

目的:探讨RNA干扰大鼠CREB结合蛋白(rCBP)基因对大鼠血管平滑肌细胞(VSMCs)增殖的影响.方法:根据CBP cDNA序列构建可同时表达4条针对rCBP mRNA特异的短发夹RNA(shR-NA)、携带绿色荧光蛋白的腺病毒(CBP-shRNA/Ad).以空腺病毒为转染对照组、含非特异性shRNA编码序列的腺病...

Inhabition of proliferation of vascular smooth muscle cells by CBP gene-silencing

Objective： To explore the effect of CREB binding protein（rCBP） gene-silencing on vascular smooth muscle cells（VSMCs） proliferation by RNA interference.Methods： According to the CBP cDNA sequence,adenovirus with green fluorescent protein was constructed which could encode specific short hairpin RNA（shRNA） of rCBP mRNA（CBP-shRNA/Ad）.Empty adenovirus was taken as transfection control group,adenovirus with non-specific shRNA coding sequence as negative control group,and CBP-shRNA/Ad with multiplicity of infection 12,16,25 and 50 as intervention groups,VSMCs 2d primed by 0.1U/mL thrombin was transfected.Then the change of CBP mRNA and protein were detected by RT-PCR and western blot;cell cycle was examined by flow cytometry to evaluate VSMCs proliferation.The adverse effect of CBP-shRNA/Ad on VSMCs was investigated by invert microscope.Results： CBP-shRNA/Ad could obviously inhibit CBP high-expression induced by thrombin at mRNA and protein level（P〈0.05） in dose-dependence.CBP-shRNA/Ad increased cells rate in G0/G1 phase（P〈0.05）and decreased cells rate in S phase（P〈0.05）via reduction of CBP content.Results of invert microscope showed cells had the good adherence ability.Conclusions： RNA interference could down regulate rCBP expression selectively,inhibit VSMCs proliferation apparently,without obvious adverse effect.