【摘要】 目的探讨慢病毒介导的RNAi靶向沉默人细胞间黏附分子1(ICAM-1)基因对人淋巴瘤Raji细胞迁移及侵袭的影响。方法常规培养Raji细胞并随机分为空白对照组、NC组及干扰组。将p LKO. 1-ICAM-1shRNAGFP载体、ps PAX2载体及p MD2. G载体共转染HEK293T细胞并包装慢病毒转染干扰组,同时构建无干扰能力的阴性对照病毒转染NC组,空白对照组不予转染。用RT-PCR法检测细胞内ICAM-1 mRNA表达,用Western blotting法检测细胞内ICAM-1蛋白表达,用细胞划痕实验检测细胞迁移能力,用Transwell实验检测细胞侵袭能力。结果干扰组ICAM-1 mRNA及蛋白相对表达量低于NC组、空白对照组(P均<0. 05),细胞迁移率低于NC组、空白对照组(P均<0. 05),细胞穿膜数低于NC组与空白对照组(P均<0. 05)。结论慢病毒介导的RNAi靶向沉默ICAM-1基因表达可有效抑制人淋巴瘤Raji细胞的迁移与侵袭。更多还原

【Abstract】 Objective To observe the effects of targeted silencing human intercellular adhesion molecule 1( ICAM-1) gene on the migration and invasion of human lymphoma Raji cells. Methods The Raji cells were cultured routinely and divided into the blank control group,NC group and interference group randomly. The p LKO. 1-ICAM-1 shRNA-GFP,ps PAX2 and p MD2. G carrier were transfected into HEK293 T cells to package with lentiviral for the interference group,we also constructed negative control virus without interfering ability for NC group and the blank group was left untreated. RTPCR was used to detect the changes of ICAM-1 mRNA expression,and Western blotting was used to detect the protein expression of ICAM-1,and meanwhile,the changes of cell migration ability were detected by cell scratch test and the changes of cell invasion ability were detected by Transwell experiment. Results The relative mRNA and protein expression levels of ICAM-1 were lower in the interference group than in the NC group and blank control group( both P < 0. 05). The cell migration rate of the interference group was lower than those of NC group and blank control group( both P < 0. 05). The number of transmembrane cells in the interference group was significantly lower than that of the NC group and blank control group( both P < 0. 05). Conclusion The lentiviral mediated RNAi targeted silencing ICAM-1 gene expression can effectively inhibit the migration and invasion of human lymphoma Raji cells.更多还原