Two essential characteristics of therapeutic nanocarriers are their rapid uptake into, and their specific recognition by tumor cells.Here we aim to develop a facile dual-ligand modification strategy to endow nanocarfier with tumor-specific penetration feature.Folate (FA) and cell-penetrating peptide (TAT peptide) were conjugated to lipid materials by simple chemical reactions using different chain lengths of polyethylene glycol (PEG) as linkers.Liposomes with an optimal ligand combination of FA and TAT peptide were constructed using a controllable method.Particle size and zeta-potential determinations indicated that long chain PEG provided enough space for masking the cationic TAT peptide, and the dual-ligand liposome had a moderate negative potential of-4.4 mV.Antitumor activity of the liposome was significantly boosted after dual-ligand modification and its IC50 value was 5.3-times and 2.6-times lower than its single-ligand counterparts.It was found that cellular uptake of the dual-ligand liposome was 2.8-times and 1.6-times faster than that of its single-ligand counterparts.Interestingly, recognition and binding of FA to FA receptors activated translocation ability of the TAT peptide.Mechanism studies revealed that clathrin-and macropinocytosis mediated endocytosis contributed to cell entry of the dual-ligand liposome.As expected, dual-ligand modification increased accumulation and prolonged retention of the liposome in tumors.Collectively, this study demonstrates the feasibility and great potential of this dual-ligand modification strategy for achieving efficient tumor-targeted drug delivery.