聚甲基丙烯酸甲酯纳米颗粒作为A549细胞中Survivin mRNA特异性分子信标载体

B. Adinolfi, F. Baldini, A. Giannetti, S. Tombelli, C. Trono, G. Sotgiu, G. Varchi, M. Pellegrino, C. Domenici
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引用次数: 0

摘要

使用反义寡核苷酸分子信标,当它们与目标mRNA杂交时能够产生荧光信号,代表了癌症治疗的一种创新策略。这种方法能够将感知特定mRNA的能力与药物沉默活性结合起来,防止与癌症发展相关的蛋白质过度表达。在这种情况下,该策略通过使用有效的递送系统,将主要针对肿瘤细胞的治疗的非特异性毒性降至最低。本研究的目的是研究聚甲基丙烯酸甲酯纳米颗粒(PMMA-NPs)在A549人肺腺癌上皮细胞中作为靶向survivin mRNA的寡核苷酸分子信标(MB)载体的能力。此外,本文还重点讨论了在体外实验中需要适当的健康对照。特别是,我们首先在溶液中对survivin-MB进行了表征,以验证其功能,然后通过共聚焦显微镜在A549细胞中验证了PMMA-NPs促进MB内化的能力。以成人合流人真皮成纤维细胞(HDFa)作为健康对照。结果表明,PMMA-NPs促进了survivin-MB的细胞摄取,使用10µg/mL PMMA-NPs作为survivin-MB的载体1h - 30min可能是一种有希望的减少癌细胞增殖的策略,同时避免对健康细胞产生可检测到的后果。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Polymethylmethacrylate Nanoparticles as Vehicle for a Molecular Beacon Specific for Survivin mRNA in A549 Cells
The use of antisense oligonucleotide molecular beacons, able to generate a fluorescent signal when they hybridize with their target mRNA, represents an innovative strategy in cancer. This approach is able to conjugate the ability of sensing specific mRNA with the pharmacological silencing activity, preventing the overexpression of proteins associated to cancer development. In this context, this strategy minimizes the non-specific toxicity addressing the therapy mainly towards the tumor cells by using effective delivery systems. The aim of this work was to investigate the ability of polymethylmethacrylate nanoparticles (PMMA-NPs) to act as vehicle of an oligonucleotide molecular beacon (MB) targeting survivin mRNA in A549 human lung adenocarcinoma epithelial cells. Furthermore, this paper focuses the attention on the need for having an appropriate healthy control in in-vitro experiments. In particular, the survivin-MB was firstly characterized in solution in order to verify its functionality and then the PMMA-NPs ability to promote the MB internalization was verified in A549 cells by confocal microscopy. Confluent Human Dermal Fibroblasts from adult (HDFa) were used as healthy control. The results showed that PMMA-NPs promote the survivin-MB cellular up-take and that the use of 10 µg/mL PMMA-NPs as carrier for survivin-MB for 1h 30 min might be a promising strategy to reduce cancer cell proliferation avoiding detectable consequences on the healthy cells.
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