A new Ag(I) complex (A 3) was synthesized and evaluated for its anticancer activity against human cancer cell lines. Materials and Methods: The complex A 3 was characterized by 1 H, 13 C, and 31 P nuclear magnetic resonance (NMR), infrared (IR) spectra, elemental analysis, and X-ray crystallography. The interaction of the complex with CT-DNA was studied by electronic absorption spectra, fluorescence spectroscopy, and cyclic voltammetry; cell viability (%) was assessed by absorbance measurement of the samples. Results: The interaction mode of the complex A 3 with DNA is electrostatic, and this complex shows good potential in anticancer properties against HCT 116 (human colorectal cancer cells) and MDA-MB-231 (MD Anderson-metastatic breast) cell lines with 0.5 micromolar concentrations. Conclusion: The Ag(I) complex could interact with DNA noncovalently and has anticancer properties.