The synthesis, characterization, DNA interaction and antiproliferative behavior of new π-arene ruthenium(II) piano-stool complexes with nitrogen ligands are described. Three series of organometallic compounds of formulae [RuCl(2)(η(6)-p-cym)L] were synthesized (with L=2-, 3- or 4-methylpyridine; L=2,3-, 2,4-, 2,5-, 3,4-, 3,5-dimethylpyridine and L=1,2-, 1,3- 1,4-methylaminobenzene). The crystal structures of [RuCl(2)(p-cym)(4-methylpyridine)], [RuCl(2)(p-cym)(3,4-dimethylpyridine)] and [RuCl(2)(p-cym)(1,4-methylaminobenzene)] were resolved and the characterization was completed by spectroscopic UV-vis, FT-IR and (1)H NMR studies. Electrochemical experiments were performed by cyclic voltammetry to estimate the redox potential of the Ru(II)/Ru(III) couple. The interaction with plasmid pBR322 DNA was studied through the examination of the electrophoretical mobility and atomic force microscopy, and interaction with ct-DNA by circular dichroism, viscosity measurements and fluorescence studies based on the DNA-ethidium bromide complex. The antiproliferative behavior of the series with L=methylpyridine was assayed against two tumor cell lines, i.e. LoVo and MiaPaca. The results revealed a moderate cytotoxicity with a higher activity for the LoVo cell line compared to the MiaPaca one.