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为研究反义单核细胞趋化蛋白－１ 转基因表达对单核细胞进入动脉壁的作用，首先构建了表达反义单核细胞趋化蛋白－ １ 基因的逆转录病毒重组体，并观察它在培养的细胞中的表达。将家兔单核细胞趋化蛋白－ １ ｃＤＮＡ 反向插入到ｐＬＮＣＸ，构成ＬＮＣＸ－ａｎｔｉ－ ＭＣＰ－１ 重组病毒质粒。再将重组质粒转染φ－２ 细胞，继以φ－ ２ 细胞产生的病毒上清感染ＰＡ３１７细胞，取得Ｇ４１８ＰＡ３１７ 抗细胞克隆。上述细胞经扩增培养，收集病毒上清并感染ＮＩＨ３Ｔ３ 细胞后进行检测。结果发现，病毒的滴度为５．６×１０７ ＣＦＵＬ，感染的ＮＩＨ３Ｔ３ 细胞中有重组病毒的整合。重组病毒感染培养的家兔动脉平滑肌细胞后，用聚合酶链反应检测发现，感染的平滑肌细胞基因组ＤＮＡ中有重组病毒整合；ＲＮＡｓｌｏｔ 杂交结果显示，感染的平滑肌细胞中有反义单核细胞趋化蛋白－１ 的表达，与未感染的平滑肌细胞相比，感染的平滑肌细胞中单核细胞趋化蛋白－ １ ｍＲＮＡ 的表达明显受到抑制。结果提示，反义单核细胞趋化蛋白－ １ 逆转录病毒表达载体在培养的动脉平滑肌中能表达反义基因并抑制靶基因的表达，为进一步开展体内实验研究奠定了基础
In order to study the effect of antisense monocyte chemoattractant protein-1 transgene on monocyte entry into the arterial wall, a recombinant retrovirus expressing antisense monocyte chemoattractant protein-1 gene was constructed and observed Expression in cultured cells. Rabbit monocyte chemotactic protein - 1 cDNA was reverse inserted into pLNCX to construct LNCX-anti-MCP-1 recombinant virus plasmid. The recombinant plasmids were transfected into φ-2 cells, followed by infecting PA317 cells with virus supernatant from φ-2 cells to obtain G418PA317 anti-cell clone. After the above-mentioned cells were expanded and cultured, the virus supernatant was collected and infected with NIH3T3 cells for detection. As a result, it was found that the titer of the virus was 5.6 × 10 7 CFU × L, and there was integration of the recombinant virus in the infected NIH3T3 cells. Recombinant virus infection in cultured rabbit arterial smooth muscle cells, detected by polymerase chain reaction, the infected infected smooth muscle cell genome DNA recombinant virus integration; RNAslot hybridization results showed that infected smooth muscle cells with antisense monocyte chemotactic The expression of MCP - 1 mRNA was significantly inhibited in infected VSMCs compared with uninfected smooth muscle cells. The results suggest that antisense monocyte chemoattractant protein - 1 retroviral vector can express antisense gene and inhibit target gene expression in cultured arterial smooth muscle, which lays the foundation for further in vivo experimental research