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目的:探讨聚乙烯亚胺-壳聚糖(PEI-CS)/si RNA复合颗粒对肝癌耐药细胞BEL7402/5-FU中MRE11表达的影响。方法:采用复凝聚法将PEI-CS(100μg/m L)与不同浓度的MRE11 si RNA-FAM形成PEI-CS/si RNA复合颗粒,并转染BEL7402/5-FU细胞,用荧光显微镜和Real-time PCR检测转染效率和沉默效率。结果:荧光显微镜观察结果显示:转染细胞48 h后,3.125、6.25、12.5、25、50μg/m L的si RNA与PEI-CS形成的复合颗粒的转染率分别为62.31%、76.09%、79.99%、86.49%、96.59%。转染细胞48、72、96 h后,12.5μg/m L的si RNA与PEI-CS形成的复合颗粒的转染率分别为78.22%、55.76%、42.85%,25μg/m L的si RNA与PEI-CS形成的复合颗粒的转染率分别为83.67%、74.23%、67.45%。Real-time PCR检测结果显示:25μg/m L的si RNA与PEI-CS形成的复合颗粒转染48小时后,对BEL7402/5-FU细胞中MRE11基因的沉默效率为35.4%。结论:聚乙烯亚胺-壳聚糖/si RAN复合颗粒能有效转染肝癌耐药细胞Bel7402/5-FU,并对BEL7402/5-FU细胞中MRE11基因表达有一定抑制作用。
Objective: To investigate the effect of polyethyleneimine-chitosan (PEI-CS) / si RNA composite particles on the expression of MRE11 in drug-resistant hepatocarcinoma cell line BEL7402 / 5-FU. METHODS: PEI-CS (100μg / ml) and different concentrations of MRE11 si RNA-FAM were used to form PEI-CS / si RNA composite particles by complex coacervation and transfected into BEL7402 / 5- Transfection efficiency and silencing efficiency were detected by time PCR. Results: The results of fluorescence microscopy showed that the transfection rates of the composite particles of si RNA and PEI-CS of 3.125, 6.25, 12.5, 25 and 50 μg / m L were 62.31% and 76.09%, respectively, 48 h after transfection, 79.99%, 86.49%, 96.59%. After 48, 72 and 96 h of transfection, the transfection rates of 12.5 μg / ml L siRNA and PEI-CS composite particles were 78.22%, 55.76%, 42.85% and 25 μg / ml respectively The transfection rates of composite particles formed by PEI-CS were 83.67%, 74.23% and 67.45%, respectively. Real-time PCR results showed that the silencing efficiency of MRE11 gene in BEL7402 / 5-FU cells was 35.4% after transfected with 25μg / ml siRNA and PEI-CS for 48 hours. CONCLUSION: Polyethylenimine-chitosan / si RAN composite particles can effectively transfect Bel7402 / 5-FU cells and inhibit the expression of MRE11 gene in BEL7402 / 5-FU cells.