文献：Design of pH-sensitive methotrexate prodrug-targeted curcumin nanoparticles for efficient dual-drug delivery and combination cancer therapy

文献链接：https://www.tandfonline.com/doi/full/10.2147/IJN.S152312#d1e205

作者：Jiajiang Xie,Zhongxiong Fan,Yang Li,Yinying Zhang,Fei Yu,Guanghao Su

摘要：

To investigate the intracellular drug/nanocarrier delivery of MTX-Imine-M-CUR, Cy5.5-labeled DSPE-PEG was synthesized for use as a near-infrared fluorescent probe (DSPE-PEG-Cy5.5 was conjugated between DSPE-PEG-NH2 and Cy5.5 NHS ester via amide linkage). HeLa cells were seeded into 24-well plates at a density of 1.0×104 cells per well. After culturing for 24 h, the cells were treated with MTX-Imine-MCy5.5-CUR at 37°C for predetermined incubation time periods. Afterward, the cells were washed three times with PBS and fixed with 4% (v/v) paraformaldehyde for 15 min. Finally, the cells were washed by cold PBS twice, stained with DAPI for 10 min, and observed using a Leica TCS SP5 CLSM.

为了研究MTX-Imine-M-CUR的细胞内药物/纳米载体递送，合成了Cy5.5标记的DSPE-PEG作为近红外荧光探针（DSPE-PEG-Cy5.5通过酰胺键结合在DSPE-PEG-NH2和Cy5.5 NHS酯之间）。

HeLa细胞以每孔1.0×104个细胞的密度接种到24孔板中。培养24小时后，在37°C下用MTX-Imine-MCy5.5-CUR处理细胞预定的孵育时间。然后，用PBS洗涤细胞三次，用4%（v/v）多聚甲醛固定15分钟。

最后，用冷PBS洗涤细胞两次，用DAPI染色10分钟，并使用Leica TCS SP5 CLSM观察。

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