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首页> 外文期刊>RSC Advances >Functionalized graphene oxide for anti-VEGF siRNA delivery: preparation, characterization and evaluation in vitro and in vivo
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Functionalized graphene oxide for anti-VEGF siRNA delivery: preparation, characterization and evaluation in vitro and in vivo

机译：功能化氧化石墨烯用于抗VEGF siRNA的递送：体外和体内的制备，表征和评估

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摘要

RNA interference (RNAi) treatment is a promising and effective method for gene therapy in cancer treatments. Small interference RNA (siRNA) plays an indispensable role in the process of RNAi, resulting in gene silencing. However, naked siRNA has difficulty in crossing the cell membrane and can easily be deactivated by enzymolysis. An ideal carrier is required for siRNA delivery to overcome these disadvantages. In this study, GO–PLL–SDGR (poly-L-lysine and Arg-Gly-Asp-Ser functionalized graphene oxide), a graphene oxide (GO)-based carrier that can actively target tumors, was prepared and characterized. Results of an agarose gel retardation assay indicated that 10 μg of GO–PLL–SDGR could load 1 μg of VEGF-siRNA. It was found that the release of VEGF-siRNA from GO–PLL–SDGR/VEGF-siRNA was slow and sustained. The efficiency of gene silencing and the tumor growth inhibitory activity of GO–PLL–SDGR/VEGF-siRNA were investigated both in vitro and in vivo by RT-qPCR, ELISA and an S180 tumor-bearing mice model. RT-qPCR and ELISA assays revealed that the expressions of VEGF-mRNA and VEGF protein were down-regulated by 40.86% and 51.71%, respectively, in vitro. In vivo, the Cy-3 labeled VEGF-siRNA was observed to assemble in tumor tissues, and the tumor inhibitory rate of GO–PLL–SDGR/VEGF-siRNA was 51.74%. What's more, GO–PLL–SDGR exhibited low cytotoxicity in the MTT assay. As all the evidence shows above, GO–PLL–SDGR could be used in siRNA delivery systems as a non-viral tumor targeting carrier.
机译：RNA干扰（RNAi）治疗是在癌症治疗中进行基因治疗的有前途且有效的方法。小干扰RNA（siRNA）在RNAi的过程中起着不可或缺的作用，从而导致基因沉默。但是，裸露的siRNA难以穿过细胞膜，并且很容易被酶解灭活。 siRNA递送需要理想的载体来克服这些缺点。在这项研究中，GO–PLL–SDGR（聚- L -赖氨酸和Arg-Gly-Asp-Ser功能化氧化石墨烯）是一种基于氧化石墨烯（GO）的载体，可主动靶向肿瘤准备并进行了表征。琼脂糖凝胶阻滞分析的结果表明，10μgGO–PLL–SDGR可以加载1μgVEGF-siRNA。发现GO-PLL-SDGR / VEGF-siRNA的VEGF-siRNA释放缓慢且持续。通过RT-qPCR，ELISA和体外和体内方法研究了GO–PLL–SDGR / VEGF-siRNA的基因沉默效率和肿瘤生长抑制活性。 S180荷瘤小鼠模型。 RT-qPCR和ELISA检测表明，在体外 中，VEGF-mRNA和VEGF蛋白的表达分别下调了40.86％和51.71％。 体内，观察到Cy-3标记的VEGF-siRNA在肿瘤组织中聚集，GO–PLL–SDGR / VEGF-siRNA的抑瘤率为51.74％。此外，GO–PLL–SDGR在MTT分析中显示出低细胞毒性。如以上所有证据所示，GO–PLL–SDGR可作为非病毒肿瘤靶向载体用于siRNA递送系统。

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