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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。
AIM: To investigate the effect of PECAM-1 on the proliferation of murine hemangioendothelioma (EOMA) cells and the expression of VEGF in siRNA CD31 targeting silencing of vascular endothelial cells Impact. Methods: The experiment was divided into two groups: naked siRNA CD31 group, siRNA CD31-FAM group, stable negative control group (SNC) and control group (Opti-Med). The cationic liposomes (RNAi-mate) O-methyl-modified siRNA CD31 was transfected into EOMA cells in vitro. The transfection efficiency of siRNA CD31 was observed by laser scanning confocal microscopy. The proliferation of EOMA cells was detected by MTT assay The expression of PECAM-1 and VEGF in EOMA cells were detected by RT-PCR and Western blotting respectively. Results: Compared with SNC group and blank control group, the expression of PECAM-1 mRNA and protein, VEGF mRNA and protein in naked siRNA CD31 and siRNA CD31-FAM transfected EOMA cells were significantly decreased (both P <0.01). Compared with SNC group, the inhibitory rate of EOMA cell proliferation was significantly increased in untreated siRNA CD31 group and siRNA CD31-FAM group [(18.82 ± 1.46)%, (18.91 ± 2.21)% vs (0.61 ± 1.06)%, both P <0.01] . Conclusion: The silencing of PECAM-1 gene in EOMA cells by siRNA CD31-liposome complex can inhibit the expression of VEGF mRNA and protein and thus inhibit the proliferation of EOMA cells.