Preparation of paclitaxel-loaded nanoparticles targeting liver cancer stem cells and their effects on liver cancer Huh-7 and HepG2 cells
10.3760/cma.j.cn115355-20200601-00297
- VernacularTitle:靶向肝癌干细胞的紫杉醇纳米粒的制备及其对肝癌Huh-7和HepG2细胞作用的研究
- Author:
Cheng JIN
;
Shuangquan WANG
;
Ling BAI
- From:
Cancer Research and Clinic
2021;33(2):99-103
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the preparation of paclitaxel-loaded nanoparticles targeting liver cancer stem cells and their effects on liver cancer HepG2 cells (CD133 positive subset accounting for 8%) and Huh-7 cells (CD133 positive subset accounting for 65%).Methods:Poly (lactic-co-glycolic acid)-loaded paclitaxel nanoparticles were prepared by using emulsification-solvent evaporation method. Paclitaxel-loaded nanoparticles decorated with anti-CD133 antibody, called targeted nanoparticles, were prepared by using 1- (3-dimethylaminopropyl)-3-ethylcarbodiimide/N-hydroxysuccinimide (EDC/NHS) cross-linking method. The manifestations and physicochemical characteristics of the nanoparticles including encapsulation efficiency, loading efficiency, particle size distribution, morphology and release in vitro were studied. Liver cancer Huh-7 and HepG2 cells accompanying with paclitaxel-loaded nanoparticles or targeted nanoparticles were cocultured. The uptake and accumulation of nanoparticles by liver cancer cells were analyzed by using flow cytometry, and positive cell proportion of CD133 was also detected. Cell survival was analyzed by using plate clonogenesis assay.Results:Scan electromicroscopy result showed particle size of targeted nanoparticles was (429.26±41.53) nm with zeta potential of -11.2 mV; targeted nanoparticles were possessed with spherical morphology and higher encapsulation efficiency [(87.53±5.90) %]. Flow cytometry showed that in Huh-7 cells at 37℃, the fluorescence intensity of targeted nanoparticles group (13 397±720) was higher than that of paclitaxel-loaded nanoparticles group (6 898±604), and the difference was statistically significant ( P < 0.05); there was no statistically difference in the fluorescence intensity of HepG2 cells in paclitaxel-loaded nanoparticles group and targeted nanoparticles group at 37 ℃ (7 899±343 vs. 8 432±516, P>0.05). CD133 positive cell proportion of Huh-7 cells in targeted nanoparticles group [(15.7±2.6)%] was lower than that in paclitaxel-loaded nanoparticles group [(54.9±7.4)%], and the difference was statistically significant ( t = 7.31, P = 0.008); there was no statistical difference of HepG2 cells between the two goups ( P > 0.05). Plate clonogenesis assay showed that the cell survival rate of Huh-7 cells in targeted nanoparticles group was lower than that in paclitaxel-loaded nanoparticles group at different time points, and the difference was statistically significant ( F = 5.56, P = 0.009); but there was no statistically significant difference in HepG2 cell survival rate between the two groups ( F = 1. 19, P = 0.142). Conclusion:Prepared nanoparticles targeting liver cancer stem cells have a good inhibitory effect on liver cancer Huh-7 cells.
