ORC1基因RNA干扰对大鼠血管平滑肌细胞表型的影响  

Effect of RNA targeting ORC1 gene on phenotype of rat vascular smooth muscle cells

作　　者：李建涛[1] 舒茂琴[1] 冯媛媛[1] 

机构地区：[1]第三军医大学西南医院心血管内科,重庆市介入心脏病学研究所,重庆400038

出　　处：《第三军医大学学报》2009年第6期491-494,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30470727)~~

摘　　要：目的以RNA干扰抑制血管平滑肌细胞(vascular smooth muscle cells,VSMCs)ORC1基因,探讨ORC1基因表达抑制后对VSMCs表型的影响。方法实验设置正常对照组、阴性siRNA组及阳性siRNA组。应用Western blot检测ORC1基因表达的变化;免疫荧光染色后激光共聚焦显微镜观察细胞形态结构;应用流式细胞仪检测细胞表型标志蛋白表达。结果①siRNA转染后,阳性siRNA组ORC1基因表达水显著降低,而正常对照组及阴性siRNA组间ORC1基因表达水平无显著差异。②siRNA转染后,阳性转染组细胞呈现收缩型形态特征,空白对照组及阴性对照组细胞呈现合成型形态特征。③siRNA转染使ORC1表达减弱后,VSMCs收缩型标志物α-平滑肌肌动蛋白(α-SM actin)和平滑肌肌动蛋白重链2(SM-2)表达水平明显升高,合成型标志物骨桥蛋白(osteopontin)表达水平明显降低,空白对照组及阴性对照组间3种标志物表达水平无显著差异。结论RNA干扰介导的ORC1基因沉寂可促使VSMCs由合成型向收缩型转化。Objective To investigate the influence of RNA interference targeting ORC1 gene on the phenotype of rat vascular smooth muscle cells （VSMCs）. Methods VSMCs were transfected with siRNA targeting ORC1 gene by liposome. The expression of ORC1 protein was detected by Western blotting. The changes of phenotype-dependent markers was analyzed by flow cytometry. Results After transfected of siRNA targeting ORC1 genes, the expression of ORC1 was lower than that in the control groups （non-transfection and negative siRNA）. ORC1 gene silencing increased the expression of VSMC contractile markers oL-SM actin and SM-2, and decreased the expression of synthetic marker osteopontin. ORCI gene silencing greatly affected VSMC differentiated morphology. Conclusion ORC1 gene silenced by RNA interference can mediate the transition of VSMCs from synthetic phenotype to contractile phenotype.

关 键 词：ORC1 RNA干扰 血管平滑肌细胞 表型 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]