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Folic acid-modified carboxymethyl chitosan was polymerized withN-3-acrylamidophenylboronic acid monomer to prepare tumor-targeting nanoparticles (NPs). Nanoparticles’ size was characterized by dynamic light scattering, while the micromorphology was observed through transmission electron microscopy (TEM) and scanning electronic microscope (SEM). Doxorubicin was then loaded into prepared nanoparticles. The cellular uptake of these NPs was investigated in monolayer cell model and multicellular spheroids. The results revealed that FA-modified nanoparticles possess excellent ability of accumulation and penetration in 2D and 3D cell models. It was also found that these nanoparticles enhanced drug accumulation in tumor, which finally increased antitumor efficiency in H22 tumor-bearing mice. [ABSTRACT FROM PUBLISHER]