Fernandes Alexandra R, Baptista Pedro V
UCIBIO, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus da Caparica, 2829-516, Caparica, Portugal.
Methods Mol Biol. 2017;1530:319-336. doi: 10.1007/978-1-4939-6646-2_19.
Multifunctionalized gold nanobeacons (Au-nanobeacon) combine, in a single and unique platform, targeting, detection and silencing providing an effective impact in clinics boosting cancer theranostics. Here, we describe a nano-integrated platform based on Au-nanobeacons able to detect and inhibit gene expression specifically in cancer cells. The surfaces of gold nanoparticles (AuNPs) are functionalized with targeting peptides to enhance tumor cell recognition and uptake, and with fluorescently labeled antisense DNA hairpin oligonucleotides to detect AuNPs. These oligonucleotides, upon recognition and hybridization to the target, open their structure resulting in separating apart the dye and the quencher allowing the fluorophore to emit light and to monitor the intracellular interactions of AuNPs with the target and the specific silencing of gene expression. This strategy allows inhibiting KRAS gene expression in colorectal carcinoma cell lines with no relevant toxicity for healthy fibroblasts. Importantly, this nano-integrated platform can be easily adapted to hybridize with any specific target thus providing real benefits for the diagnosis and treatment of cancer.
多功能金纳米信标（Au - 纳米信标）在一个独特的单一平台上结合了靶向、检测和沉默功能，对临床癌症诊疗产生有效影响。在此，我们描述了一种基于Au - 纳米信标的纳米集成平台，该平台能够在癌细胞中特异性地检测和抑制基因表达。金纳米颗粒（AuNP）的表面用靶向肽进行功能化修饰，以增强肿瘤细胞的识别和摄取，并与荧光标记的反义DNA发夹寡核苷酸结合以检测AuNP。这些寡核苷酸在识别并与靶标杂交后，会打开其结构，导致染料和猝灭剂分离，使荧光团发光，并监测AuNP与靶标的细胞内相互作用以及基因表达的特异性沉默。该策略能够在结肠癌细胞系中抑制KRAS基因表达，而对健康成纤维细胞无明显毒性。重要的是，这种纳米集成平台可以轻松地与任何特定靶标杂交，从而为癌症的诊断和治疗带来实际益处。
