The combination of selenium and polysaccharides is one of the significant ways to ameliorate the anti-cancer effects of polysaccharides. PLP50-1, a homogeneous polysaccharide purified from the aqueous extract of Paeonia lactiflora, had a molecular weight of 1.52 × 10 Da and consisted of α-D-Glcp-(1→, →4)-α-D-Glcp-(1→, →6)-α-D-Glcp-(1→, →4,6)-α-D-Glcp-(1→, and →6)-β-D-Fruf-(2→. PLP50-1 showed weak anti-tumor effects against A549 cells. To ameliorate the activity of PLP50-1, the complex nanoparticles combining P. lactiflora polysaccharide with selenium were constructed successfully. Structural properties of the polysaccharide-based selenium nanoparticles (PLP-SeNPs) were clarified using various means. The results displayed that a kind of monodisperse spherical nanoparticles containing high selenium content (39.1 %) with controllable size was constructed and showed satisfactory stability. The cellular anti-tumor assay indicated that PLP-SeNPs had stronger antiproliferative activity against A549 cells than PLP50-1. Additionally, the zebrafish experiments displayed that PLP-SeNPs inhibited the proliferation and migration of A549 cells significantly and blocked the angiogenesis.