Glioma is one of the most deadly tumors due to invasive growth. The treatment effect is poor due to the presence of the blood brain barrier (BBB) and blood tumor barrier (BTB) and insufficient drug targeting. DNA tetrahedron (TDN) is considered to have great potential for drug delivery and maybe a novel therapeutic strategy for glioma. In this study, we have developed a doxorubicin loaded-DNA tetrahedron modified by a Gint4.T aptamer([email protected]) that could target PDGFRβ for therapy of glioma. The TDN and Apt-TDN were self-assembled by one-step synthesis. The characterization and stability were detected using gel electrophoresis analysis, dynamic light scatting and atomic force microscopy. The cytotoxicity of TDN in vitro was determined by CCK-8 assays. Fluorescence spectrophotometry was used to measure the drug loading capacity. Fluorescence microscopy imaging and flow cytometry were used to survey the cellular uptake. The cell viability, cell cycle and early apoptosis were estimated when the U87MG cells were treated with DOX, [email protected] and [email protected] The rusults showed that a single Apt-TDN could carry 60 doxorubicin molecules. The 3D structure of the DNA tetrahedron remarkedly enhanced drug stability in fetal bovine serum, remaining intact for 7h leastwise. The CCK-8 assay showed that the activity of the U87 cells was not affected after co-cultured with different concentrations(10–500 nM) of TDN for 24 h and 48 h. The aptamer Gint4.T enhanced the targeted cellular uptake of TDN by U87MG cells. Dox-loaded Apt-TDN inhibited cell viability and induced early apoptosis. In conclusion this study establishes that the Gint4.T-modified DNA tetrahedron could target PDGFRβ and provide a novel therapy with promising clinical application for gliomas.