Brazilian Research Agencies: FAPEMIG, FAPESB, FAPESP, CNPq and CAPES. D.E.L.C. thanks FAPEMIG for a fellowship, and K.M.O. is supported by a postdoctoral fellowship grant from CAPES (PNPD program). Also, R.S.C. would like to thank the financial support provided by PROPP/ UFOP, FAPEMIG (APQ-01674-18), and CNPq (grants 403588/2016-2 and 308370/2017-1). The authors thank the IFSC-USP (E.E. Castellano and J.A. Ellena) for X-ray crystallography measurements and the “Laboratório Multiusuário de Caracterização de Moléculas”/UFOP for the NMR facilities.Two new Ru(II)-based complexes containing 2-thiouracil derivatives, known as 2-thiouracil (2TU) and 6-methyl-2-thiouracil (6m2TU), were synthesized using cis,trans-[RuCl2(PPh3)2(bipy)] as a precursor. The obtained compounds with a general formula trans-[Ru(2TU)(PPh3)2(bipy)]PF6 (1) and trans-[Ru(6m2TU)(PPh3)2(bipy)]PF6 (2) were characterized by analytical techniques such as NMR, UV-vis, and IR spectroscopies, elementary analysis, mass spectrometry, and single-crystal X-ray diffraction. Moreover, the investigation of the complexes-DNA interaction were carried out using spectrophotometric titrations and showed that the complexes present a weak interaction with this biomolecule. The compounds were evaluated against HL-60, K-562, HepG2, and B16-F10 cancer cells and against noncancer cells (PBMCs). The results of the biological assay revealed that complex 2 is more promising than complex 1. Finally, the present study suggests that complexes 1 and 2 causes cell death by apoptosis, significantly increasing the percentage of apoptotic HL-60 cells, in which the compounds altered the cell cycle, reducing the cells in G1/G0, G2/M, and S phases.