Shaat Hanan, Mostafa Amany, Moustafa Moustafa, Gamal-Eldeen Amira, Emam Ahmed, El-Hussieny Enas, Elhefnawi Mahmoud
Chemistry Department, Faculty of Science, Benha University, Benha, Egypt; Nanomedicine and Tissue Engineering Laboratory, Medical Research Centre of excellence, National Research Centre (NRC), Cairo, Egypt.
Nanomedicine and Tissue Engineering Laboratory, Medical Research Centre of excellence, National Research Centre (NRC), Cairo, Egypt; Ceramics Department, NRC, Dokki, Cairo, Egypt,.
Int J Pharm. 2016 May 17;504(1-2):125-33. doi: 10.1016/j.ijpharm.2016.03.051. Epub 2016 Mar 29.
To overcome the rapid enzymatic degradation and low transfection efficiency of siRNA, the delivery carriers for siRNA is a therapeutic demand to increase its stability. Gold nanoparticles (AuNPs) modified by branched polyethyleneimine (bPEI) were developed as an efficient and safe intracellular delivery carriers for siRNA. The current study implied that siRNA designed against an oncogene c-Myc could be delivered by a modified AuNPs complex without significant cytotoxicity. The comparative semi-quantitative and quantitative real time PCR were used to measure the c-Myc gene expression after transfection with naked siRNA and siRNA/bPEI/AuNPs, but AuNPs interfered with PCR. However, the c-Myc protein translation was successfully detected in the transfected HuH7 cells with naked siRNA and siRNA/bPEI/AuNPs and it was found to be inhibited by siRNA/bPEI/AuNPs more than naked siRNA. The results validate the successful silencing of c-Myc gene. Accordingly, it may confirm the promising and effective delivery of siRNA by bPEI/AuNPs. The complex enhances the cellular uptake of siRNA without significant cytotoxicity and confirms that bPEI modified AuNPs could be used as a good candidate for safe cellular delivery of siRNA.
为克服小干扰RNA(siRNA)快速的酶促降解和低转染效率问题,siRNA的递送载体成为提高其稳定性的治疗需求。由支化聚乙烯亚胺(bPEI)修饰的金纳米颗粒(AuNPs)被开发为一种用于siRNA的高效且安全的细胞内递送载体。当前研究表明,针对癌基因c-Myc设计的siRNA可通过修饰的AuNPs复合物递送,且无明显细胞毒性。使用比较半定量和定量实时PCR来测量用裸siRNA和siRNA/bPEI/AuNPs转染后c-Myc基因的表达,但AuNPs干扰了PCR。然而,在用裸siRNA和siRNA/bPEI/AuNPs转染的HuH7细胞中成功检测到了c-Myc蛋白翻译,并且发现siRNA/bPEI/AuNPs对其的抑制作用比裸siRNA更强。结果验证了c-Myc基因的成功沉默。因此,这可能证实了bPEI/AuNPs对siRNA有前景且有效的递送作用。该复合物增强了siRNA的细胞摄取,且无明显细胞毒性,并证实bPEI修饰的AuNPs可作为siRNA安全细胞递送的良好候选物。