文献：Engineering of hybrid anticancer drug-loaded polymeric nanoparticles delivery system for the treatment and care of lung cancer therapy

文献链接：https://pubmed.ncbi.nlm.nih.gov/34282705/

作者：Yang Zhao, Kefeng Liu , Jie Li, Juan Liao,Li Ma

相关产品：

二甲亚砜（DMSO）

Hoechst 33258

Annexinv-荧光素异硫氰酸酯（FITC）

原文摘要：Chemotherapy with combination drugs has become one of the most commonly used cancer prevention treatments, with positive clinical results. The goal of this study was to develop compostable polymeric nanomaterials (NMs) for the delivery of puerarin (PRN) and 5-fluorouracil (5FU), as well as to investigate the anticancer activity of the drug delivery system (PRN-5FU NMs) against in vitro and in vivo lung cancer cells. Since double antitumor drugs PRN and 5FU are insufficiently compressed in polymer-based bio-degradable nanoparticles, encapsulation of PRN and 5FU antitumor drugs were coencapsulated with polyethylene glycol and polylactidecoglycolide nanoparticles (NMs) is efficient. The arrangement of PRN NMs, 5FU NMs, and PRN-5FU NMs, as well as the nanoparticles shape and scale, were studied using transmission electron microscopy (TEM). 5FU-PRN NMs triggered apoptosis in lung carcinoma cell lines such as HEL-299 and A549 in vitro. Acridine orange/ethidium bromide (AO/EB) and nuclear damaging staining techniques were used to observe morphologies and cell death. The mechanistic analysis of apoptosis was also confirmed by flow cytometry analysis using dual staining. When compared to free anticancer products, the hemolysis analysis findings of the 5FU-PRN NMs showed excellent biocompatibility. Taken together the advantages, this combination drug conveyance strategy exposed that 5FU-PRN NMs could have a significant promising to improve the effectiveness of lung cancer cells.

Hoechst 33258是一种可以穿透细胞膜的蓝色荧光染料，常用于细胞核的染色。它通过与DNA双链中的小沟结合，实现对细胞核的标记。除了细胞核染色外，Hoechst 33258还可用于常规的DNA染色，帮助科学家研究DNA的结构和功能。Hoechst 33258应储存在-20°C或更低温度下，并避光保存。在配制和使用过程中，也应注意避免光照和高温。Hoechst 33258在检查形态特征中的应用主要体现在细胞核的形态观察上，以下是对其应用的具体说明：

图为：Hoechst 33258结构式

Hoechst 33258在检查形态特征中的应用：

经Hoechst 33258核染色后，FUNMs、PRN NMs和PRN-5FUNMs处理的细胞比对照组细胞显示出更亮的蓝光，提示细胞核固缩和深染色。根据AO/EB染色，5FU NMs、FUNMs、PRN NMs、PRN-5FUNMs处理的HEL-299和A549细胞中有一些释放出橙色和红色荧光，比对照组（A-C）小。用AO/EB和Hoechst 33258对细胞形态进行染色，证实5FU NMs、PRN NMs和5FUPRN NMs通过诱导细胞Apoptosis 对HEL-299和A549细胞的影响。AO/EB染色可通过荧光上的差异区分Apoptosis 细胞和坏死细胞。随着细胞暴露于不同数量的5FU NMs、PRN NMs和PRN-5FU NMs中，细胞数量逐渐减少，一些细胞发出明亮的绿色和红色荧光。

图为：合成的5FU NMs、PRN NMs和PRN-5FU NMs的体外增殖结果

结论：Hoechst 33258 是一种 DNA 特异性荧光染料，它能够与细胞核中的 DNA 双螺旋的小沟区域结合。在荧光显微镜下，通过观察被 Hoechst 33258 染色的细胞核，可以清晰地看到细胞核的大小、形状、数量等形态特征。例如，在正常细胞中，细胞核通常呈现规则的圆形或椭圆形，并且大小相对一致。而在cancer细胞中，细胞核的形态可能会发生变化，如出现核增大、核形状不规则（如出现凹陷、分叶等）、多核现象等。通过用 Hoechst 33258 染色后观察，可以对细胞是否发生cancer变等异常情况进行初步判断。