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The aim of our study was to investigate DNA-binding and cytotoxic activity of the four new Ru(II) polypyridyl complexes [Ru(dmb) 2 (HMHPIP)](ClO 4 ) 2 ( 1 ), [Ru(bpy) 2 (HMHPIP)](ClO 4 ) 2 ( 2 ), [Ru(phen) 2 (HMHPIP)](ClO 4 ) 2 ( 3 ) and [Ru(dmp) 2 (HMHPIP)](ClO 4 ) 2 ( 4 ). The complexes interact with DNA through intercalative mode and show relatively high cytotoxic activity against A549 cells, no cytotoxicity toward MG-63 cells. Complexes 1 – 4 can enhance the levels of ROS in A549 cells and induce the decrease of the mitochondrial membrane potential. These complexes inhibit the cell growth in A549 cells at G0/G1 or S phase. Complex 3 activated caspase 7, and down-regulated the expression of the anti-apoptotic protein Bcl-2. Complexes 1 – 4 induce apoptosis in A549 cells through ROS-mediated mitochondrial dysfunction pathway. [ABSTRACT FROM AUTHOR]