文章：

磁性纳米颗粒介导的基因治疗诱导乳腺癌Fas凋亡通路

Magnetic nanoparticle-mediated gene therapy to induce Fas apoptosis pathway in breast cancer 

原文发布日期：2018-03-29

英文摘要：

CD95 (Fas) is a complex integral protein that can be expressed in many cells. It induces apoptosis when interacted with its ligand CD95L (FasL). However, cancer cells are resistant to CD95-induced apoptosis because of the changes in death domain (DD) of CD95 (procaspase-8 and c-Flip). In this study, magnetic nanoparticles and lipid-based gene transfection methods were performed to provide active Fas expression in breast cancer cells. Plasmid DNA (pDNA), which can express both human Fas and GFP, was transfected to MCF-7 breast cancer cells. Expression of c-FLIP and caspase-8 and effect of monoclonal antibody FasL for apoptosis stimulation were investigated. Also transfection success of methods and effects on surface protein were compared. Western blot results indicated that MCF-7 cells do not express caspase-8 but express large amount of c-FLIPL. Both lipid-based and magnetic nanoparticle-mediated gene transfection methods successfully applied. Caspase-8 apoptosis pathway was activated on transfected cells. Magnetic nanoparticle-mediated gene transfer is a successful non-viral method for transfection, and it does not affect the expression of other cell proteins, such as beta actin and lamin-B1. The raised c-FLIPL concentration in cytosol inhibits apoptosis. However, transfection of CD95-GFP-tagged pDNA significantly increases apoptosis by activating caspase-8 pathway. FasL interaction indicated a slight increase of apoptosis in the transfected cells. The method and pDNA applied in this study have potentials to be used in gene therapy for breast cancer. 

摘要翻译： 

CD95（Fas）是一种复杂的整合蛋白，可在多种细胞中表达。当其与配体CD95L（FasL）相互作用时，会诱导细胞凋亡。然而，由于CD95死亡结构域（DD）中procaspase-8和c-Flip的变异，癌细胞对CD95介导的凋亡具有抵抗性。本研究采用磁性纳米颗粒和脂质体基因转染方法，在乳腺癌细胞中实现活性Fas蛋白的表达。将可同时表达人源Fas与绿色荧光蛋白（GFP）的质粒DNA（pDNA）转染至MCF-7乳腺癌细胞系，并探究c-FLIP与caspase-8的表达情况以及单克隆抗体FasL对凋亡的诱导作用，同时比较不同转染方法的效率及对细胞表面蛋白的影响。蛋白质印迹结果表明：MCF-7细胞不表达caspase-8但高表达c-FLIPL；脂质体与磁性纳米颗粒介导的基因转染均成功实施；转染细胞中caspase-8凋亡通路被激活。磁性纳米颗粒介导的基因转染是一种成功的非病毒转染方法，且不影响β-肌动蛋白和核纤层蛋白B1等其他细胞蛋白的表达。胞质内升高的c-FLIPL浓度会抑制凋亡，但转染CD95-GFP标记的pDNA可通过激活caspase-8通路显著增强凋亡效应。FasL相互作用在转染细胞中仅引起轻微凋亡增加。本研究所采用的转染方法与pDNA在乳腺癌基因治疗领域具有应用潜力。

原文链接：

Magnetic nanoparticle-mediated gene therapy to induce Fas apoptosis pathway in breast cancer