siRNA CD31沉默PECAM-1对鼠源血管内皮瘤细胞VEGF表达的影响被引量：2

Effects of siRNA CD31-mediated PECAM-1 gene silencing on proliferation and VEGF expression in vascular endothelial cells

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摘要目的:研究siRNA CD31靶向沉默血管内皮细胞中血小板内皮细胞黏附分子1(platelet endothelial cell adhesion molecule 1,PECAM-1)基因对鼠源性血管内皮瘤(murine hemangioendothelioma,EOMA)细胞增殖及其VEGF表达的影响。方法:实验分为裸siRNA CD31组、siRNA CD31-FAM组、稳定阴性对照(SNC)组、空白对照(Opti-Med)组,以阳离子脂质体(RNAi-mate)为载体将化学合成的2'-O-甲基修饰的siRNA CD31转染体外培养的EOMA细胞,以激光共聚焦显微镜观察siRNA CD31的转染效果,以四甲基偶氮唑蓝(MTT)法检测siRNA CD31对EOMA细胞增殖的影响,以RT-PCR、Western blotting分别检测EOMA细胞中PECAM-1、VEGF的表达水平。结果:与SNC组和空白对照组比较,裸siRNA CD31和siRNA CD31-FAM转染的EOMA细胞中的PECAM-1 mRNA和蛋白、VEGF mRNA和蛋白的表达量均显著降低(均P<0.01)。与SNC组比较,裸siRNA CD31组、siRNA CD31-FAM组的EOMA细胞增殖抑制率明显上升[(18.82±1.46)%、(18.91±2.21)%vs(0.61±1.06)%,均P<0.01]。结论:采用siRNA CD31-脂质体复合物沉默EOMA细胞中的PECAM-1基因可抑制VEGF mRNA和蛋白的表达,从而抑制EOMA细胞的增殖。 Objective：To investigate the effects of siRNA CD31-targeted silencing of the platelet endothelial cell adhe- sion molecule 1 （PECAM-1） or CD31 gene on VEGF expression and proliferation in endothelial cells. Methods： Murine hemangioendothelioma cells （EOMAs） were used as a model. They were transfected with naked siRNA CD31, siRNA CD31-FAM, a stable negative control （SNC） siRNA and Opti-Med as a blank control, respectively, using lipofectamin （RNAi-mate）. After transfection, cell proliferation was assessed by MTT assays. PECAM-1 and VEGF mRNA and protein levels were determined by RT-PCR and Western blotting respectively. Results： PECAM-1 mRNA and protein levels and proliferative activity were all significantly lower in EOMAs transfected with naked siRNA CD31 and siRNA CD31-FAM than in EOMAs transfected with the SNC and Opti-MEM （P 〈0.01 ）. As compared with the SNC, naked siRNA CD31 and siRNA CD31-FAM rested significantly higher rates of proliferation inhibition （P 〈 0.01 ）. Conclusion： The chemi- cally synthesized 2＇-O-methyl-siRNA CD31 may effectively silence the target gene PECAM-1 and inhibit proliferation in EOMAs, at least partially through a VEGF signaling-dependent mechanism.

作者欧阳金生陈成水李玉苹蔡畅叶民林全陈俊杰

机构地区温州医科大学附属第一医院呼吸内科

出处《中国肿瘤生物治疗杂志》 CAS CSCD 北大核心 2014年第2期160-164,共5页 Chinese Journal of Cancer Biotherapy

基金浙江省温州市科技计划资助项目(No.Y20100182)~~

关键词小分子干扰RNA 血小板内皮细胞黏附分子血管内皮细胞生长因子血管内皮瘤细胞增殖 small interference RNA （siRNA） platelet endothelial cell adhesion molecule （PECAM） vascular endo-thelial cell growth factor（VEGF） hemangioendothelioma cell proliferation

分类号 R732.2 [医药卫生—肿瘤] R730.59 [医药卫生—肿瘤]

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