Arami Sanam, Mahdavi Majid, Rashidi Mohammad Reza, Fathi Marziyeh, Hejazi Mohammad-Saeid, Samadi Nasser
Pharmaceutical Biotechnology Department, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran; Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Biology, Faculty of Natural Science, University of Tabriz, Tabriz, Iran.
Biologicals. 2016 Nov;44(6):487-496. doi: 10.1016/j.biologicals.2016.09.005. Epub 2016 Oct 3.
As a gene delivery method in breast cancer therapy, knocking down the undesired genes in the cancerous cells would be promising. Inhibitors of Apoptosis Protein (IAP) family genes are some of the genes whose responsibility is inhibition of apoptosis in cells. Silencing these genes seems to be helpful directing the tumor cells to death. siRNA sequence designed against survivin anti-apoptotic gene can play this role if carried to the cytoplasm. Here we prepared a positive charged biocompatible nano-sized particle made up of a FeO core covered respectively by polyacrylate (PA) and polyethyleneimine (PEI) layer, which could successfully deliver the siRNA into the MCF-7 cells. The particle structure was checked and having less than 50 nm diameter in size, positive charge and, safety towards MCF-7 cells besides being able to form nanoplexes with the siRNA strand helps it entering into the biologic assays part. The siRNA delivery evaluated via flowcytometry. Apoptosis induction was determined by DAPI staining. The efficiency of survivin gene knockdown was evaluated in mRNA and protein levels using Real time PCR and western blotting methods. Overall, the FeO-PA-PEI nanoparticles can deliver siRNA effectively into the cytoplasm of the MCF-7 breast cancer cells and induce apoptosis.
作为乳腺癌治疗中的一种基因传递方法，敲除癌细胞中不需要的基因可能很有前景。凋亡抑制蛋白（IAP）家族基因的抑制剂是一些负责抑制细胞凋亡的基因。沉默这些基因似乎有助于引导肿瘤细胞走向死亡。针对存活素抗凋亡基因设计的小干扰RNA（siRNA）序列如果能转运到细胞质中就能发挥这种作用。在这里，我们制备了一种带正电荷的生物相容性纳米颗粒，它由一个FeO核分别被聚丙烯酸酯（PA）和聚乙烯亚胺（PEI）层覆盖组成，该颗粒能够成功地将siRNA递送至MCF-7细胞。对颗粒结构进行了检查，其直径小于50nm，带正电荷，对MCF-7细胞安全，此外还能够与siRNA链形成纳米复合物，这些特性有助于其进入生物学检测部分。通过流式细胞术评估siRNA的递送情况。通过4',6-二脒基-2-苯基吲哚（DAPI）染色确定凋亡诱导情况。使用实时聚合酶链反应（PCR）和蛋白质免疫印迹法在mRNA和蛋白质水平评估存活素基因敲低的效率。总体而言，FeO-PA-PEI纳米颗粒能够有效地将siRNA递送至MCF-7乳腺癌细胞的细胞质中并诱导凋亡。
