Аннотация:Abstract Objectives Aptamer sgc8c is a short DNA sequence that can target protein tyrosine kinase 7 ( PTK 7), which was overexpressed on many tumour cells. This study aimed to fabricate a novelty DNA nanostructure drug delivery system target on PTK 7‐positive cells— CCRF ‐ CEM (human T‐cell ALL ). Methods Aptamer‐modified tetrahedron DNA was synthesized through one‐step thermal annealing process. The sgc8c‐ TDN s (s‐ TDN s) loading DOX complexes were applied to investigate the effect to PTK 7‐negative and ‐positive cells. Results When s‐ TDN : DOX acted on PTK 7‐positive and ‐negative cells respectively, the complexes exhibited specific toxic effect on PTK 7‐positive cells but not on PTK 7‐negative Ramos cells in vitro research. Conclusions In this work, we successfully constructed a PTK 7‐targeting aptamer‐guided DNA tetrahedral nanostructure (s‐ TDN ) as a drug delivery system via a facile one‐pot synthesis method. The results showed that s‐ TDN : DOX exhibited enhanced cytotoxicity against PTK 7‐positive CCRF ‐ CEM cells, with a minor effect against PTK 7‐negative Ramos cells. Hence, this functionalized TDN s drug delivery system displayed its potential application in targeting PTK 7‐positive tumour T‐cell acute lymphoblastic leukaemia.