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目的观察磁性纳米微粒介导的RNA干扰技术基因转染大肠癌Lovo细胞后对Myc基因的转录及表达的影响。方法用自行制备的葡聚糖磁性纳米微粒(DCIONP)为载体,载附含有Myc基因潜在特异性干扰序列的质粒DNA(pGenesil-1-X1、pGenesil-1-X2)、含对照引物的pGen- esil-1-HK。以及裸质粒pGenesil-1。并以单纯DCIONP、不加药组作为对照组,在外加磁场的作用下,转染体外培养的Lovo细胞。通过rt-PCR及Western blot方法分别检测转染24 h后大肠癌细胞内Myc基因的mRNA转录水平及相关蛋白的表达。结果磁性转染pGenesil-1-X2的Lovo细胞,其Myc基因的mRNA转录水平明显降低,相关蛋白的表达亦显著降低。结论磁控靶向RNA干扰技术能够发挥干扰效应,抑制Myc基因mRNA的转录以及相关蛋白的表达。
Objective To investigate the effect of magnetic nanoparticle-mediated RNA interference on the transcription and expression of Myc gene in colorectal cancer Lovo cells. Methods Plasmid DNA (pGenesil-1-X1, pGenesil-1-X2) containing the specific interference sequence of Myc gene was loaded on dextran magnetic nanoparticles (DCIONP) esil-1-HK. And naked plasmid pGenesil-1. Pure DCIONP and no drug group were used as the control group, and Lovo cells cultured in vitro were transfected under the action of applied magnetic field. The mRNA and protein levels of Myc in colorectal cancer cells were detected by rt-PCR and Western blot respectively 24 hours after transfection. Results The mRNA and protein levels of Myc gene were significantly decreased in Lovo cells transfected with pGenesil-1-X2 and the expression of related proteins was also significantly decreased. Conclusion Magnetron-targeted RNA interference can exert a disruptive effect and inhibit the transcription of Myc gene and the expression of related proteins.