Journal
JOURNAL OF INORGANIC BIOCHEMISTRY
Volume 157, Issue -, Pages 62-72Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.jinorgbio.2016.01.003
Keywords
Ruthenium(II) polypyridyl complexes; Cytotoxicity in vitro; Reactive oxygen species; Mitochondrial membrane potential; Western blot analysis
Funding
- High-Level Personnel Project of Guangdong Province
- Nature Science Fund of the Department of Science and Technology [GYFYLH201315]
- First Affiliated Hospital of Guangdong Pharmaceutical University [GYFYLH201315]
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Four new ruthenium(II) polypyridyl complexes [Ru(dmb)(2)(DQTT)](ClO4)(2) (1) (DQTT = 12-(1,4-dihydroguinoxalin-6-yl)-4,5,9,14-tetraazabenzo[b]triphenylene, dmb = 4,4'-dimethyl-2,2'-bipyridine), [Ru(bpy)(2)(DQTT)](ClO4)(2) (2) (bpy = 2,2'-bipyridine), [Ru(phen)(2)(DQTT)](ClO4)(2) (3) (phen = 1,10-phenanthroline) and [Ru(dmp)(2)(DQTT)(1)(Clo(4))(2) (4) (dmp = 2,9-dimethyl-1,10-phenanthroline) were synthesized and characterized by elemental analysis, ESI-MS, H-1 NMR and C-13 NMR The cytotoxic activity in vitro of the complexes was evaluated against human BEL-7402, A549, HeLa, HepG-2 and MG-63 cancer cell lines by MTT (3-(4,5-dimethylthiazole)-2,5-diphenyltetrazolium bromide) method. The IC50 values of complexes 1-4 against BEL-7402 cells are 31.8 +/- 1.0, 35.8 +/- 1.6, 29.0 +/- 0.8 and 25.0 +/- 0.9 mu M, respectively. The morphological apoptosis was investigated with AO/EB (acridine orange/ethidium bromide) and Hoechst 33258 staining methods. The DNA damage was assayed by comet assay. The inhibition of cell migration was evaluated by the wound healing assay. The levels of ROS (reactive oxygen species) and the changes of mitochondrial membrane potential were studied under fluorescent microscope. The percentages in the cells of apoptotic and necrotic cells and the cell cycle arrest were determined by flow cytometry. The expression of Bcl-2 family proteins was investigated by western blot analysis. The results show that the complexes induce BEL-7402 cells apoptosis through a ROS-mediated mitochondria! dysfunction pathway, which was accompanied by regulation of the expression of Bcl-2 family proteins. (C) 2016 Elsevier Inc. All rights reserved.
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