Fourteen new organotin(IV) complexes were successfully synthesized and characterized by elemental analyses, FT-IR, Multinuclear (1H, 13C and 119Sn) NMR spectroscopy, HRMS and X-ray single-crystal techniques. Crystallographic data showed the complexes (1b, 2b, 3b and 5b) were macrocyclic compounds, 4b exhibits a one-dimensional spiral chain structure with distorted trigonal bipyramidal geometry, other complexes were centrosymmetric dimer and there was a Sn2O2 four-membered ring in the middle of the molecule, respectively. In vitro anticancer activity against three human tumor cell lines NCI-H460, MCF-7 and HepG2 were studied, and the dibutyltin complex 5a is a more potent antitumor agent than other complexes and cisplatin. Cell apoptosis study of 5a with the highest activity on HepG2 cancer cell lines was investigated by flow cytometry, it was shown that the antitumor activity of 5a was related to apoptosis, and it was inhibited proliferation by blocking cells in the G2/M phase. The single cell gel electrophoreses assay results show that the 5a induces DNA damage. The 5a interact with ct-DNA by intercalating the mode of interaction. UV-visible absorption spectrometry, fluorescence competitive, viscosity measurements, and gel electrophoresis results also support the intercalative mode of interaction for the 5a with DNA.