Aptamer Combined with Fluorescent Silica Nanoparticles for Detection of Hepatoma Cells.
Aptamer Combined with Fluorescent Silica Nanoparticles for Detection of Hepatoma Cells.
复制标题
适体与荧光二氧化硅纳米颗粒结合用于肝癌细胞检测
DOI：
10.1186/s11671-017-1890-6
复制
发表时间：
2017-12
影响因子：
--
通讯作者：
Lu X
中科院分区：
文献类型：
--
作者：
Hu Z;Tan J;Lai Z;Zheng R;Zhong J;Wang Y;Li X;Yang N;Li J;Yang W;Huang Y;Zhao Y;Lu X
目的本研究的目的是开发一种简单、有效的方法来用适配体标记肝癌细胞，然后用荧光二氧化硅纳米粒子（FSNPs）进行检测。方法将链霉亲和素与反相微乳液法制备的羧基修饰异硫氰酸荧光素（FITC）掺杂的二氧化硅纳米粒子缀合。将所得的链霉亲和素共轭荧光二氧化硅纳米粒子 (SA-FSNP) 与用生物素共轭适体 TLS11a (Bio-TLS11a) 标记的肝癌细胞混合。使用流式细胞术和荧光显微镜评估纳米探针的特异性和敏感性。使用MTT法在正常人肝细胞培养物中评估其毒性，并使用免疫组织化学在裸鼠中评估其毒性。结果SA-FSNPs的大小和形状均一，其荧光特性与游离FITC染料相似。 SA-FSNP 能够以优异的特异性和良好的灵敏度检测适体标记的肝癌细胞，并且它们发出强烈的、耐光漂白的荧光信号。 SA-FSNP 在体外或体内均未表现出明显的毒性作用。 结论 生物素缀合适体和 SA-FSNP 的组合有望灵敏检测肝癌细胞，并有可能检测其他肿瘤细胞类型。
PurposeThe purpose of this study is to develop a simple, effective method to label hepatoma cells with aptamers and then detect them using fluorescent silica nanoparticles (FSNPs).MethodStreptavidin was conjugated to carboxyl-modified fluorescein isothiocyanate (FITC)-doped silica nanoparticles which were prepared by the reverse microemulsion method. The resulting streptavidin-conjugated fluorescent silica nanoparticles (SA-FSNPs) were mixed with hepatoma cells that had been labeled with biotin-conjugated aptamer TLS11a (Bio-TLS11a). The specificity and sensitivity of the nanoprobes were assessed using flow cytometry and fluorescence microscopy. Their toxicity was assessed in normal human liver cell cultures using the MTT assay, as well as in nude mice using immunohistochemistry.ResultsSA-FSNPs showed uniform size and shape, and fluorescence properties of them was similar to the free FITC dye. SA-FSNPs were able to detect aptamer-labeled hepatoma cells with excellent specificity and good sensitivity, and they emitted strong, photobleach-resistant fluorescent signal. SA-FSNPs showed no significant toxic effects in vitro or in vivo.ConclusionThe combination of biotin-conjugated aptamers and SA-FSNPs shows promise for sensitive detection of hepatoma cells, and potentially of other tumor cell types as well.