Zhang Cheng, Han Bing-Jie, Zeng Chuan-Chuan, Lai Shang-Hai, Li Wei, Tang Bing, Wan Dan, Jiang Guang-Bin, Liu Yun-Jun
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, PR China.
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, PR China.
J Inorg Biochem. 2016 Apr;157:62-72. doi: 10.1016/j.jinorgbio.2016.01.003. Epub 2016 Jan 22.
Four new ruthenium(II) polypyridyl complexes Ru(dmb)2(DQTT)2 (1) (DQTT=12-(1,4-dihydroquinoxalin-6-yl)-4,5,9,14-tetraazabenzo[b]triphenylene, dmb=4,4'-dimethyl-2,2'-bipyridine), Ru(bpy)2(DQTT)2 (2) (bpy=2,2'-bipyridine), Ru(phen)2(DQTT)2 (3) (phen=1,10-phenanthroline) and Ru(dmp)2(DQTT)2 (4) (dmp=2,9-dimethyl-1,10-phenanthroline) were synthesized and characterized by elemental analysis, ESI-MS, (1)H NMR and (13)C 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 μM, 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 mitochondrial dysfunction pathway, which was accompanied by regulation of the expression of Bcl-2 family proteins.
合成了四种新型钌（II）多吡啶配合物Ru(dmb)2(DQTT)2（1）（DQTT = 12-(1,4-二氢喹喔啉-6-基)-4,5,9,14-四氮杂苯并[b]三苯撑，dmb = 4,4'-二甲基-2,2'-联吡啶）、Ru(bpy)2(DQTT)2（2）（bpy = 2,2'-联吡啶）、Ru(phen)2(DQTT)2（3）（phen = 1,10-菲咯啉）和Ru(dmp)2(DQTT)2（4）（dmp = 2,9-二甲基-1,10-菲咯啉），并通过元素分析、电喷雾电离质谱、1H NMR和13C NMR对其进行了表征。采用MTT（3-(4,5-二甲基噻唑)-2,5-二苯基四氮唑溴盐）法评估了这些配合物对人BEL-7402、A549、HeLa、HepG-2和MG-63癌细胞系的体外细胞毒性活性。配合物1 - 4对BEL-7402细胞的IC50值分别为31.8±1.0、35.8±1.6、29.0±0.8和25.0±0.9 μM。采用AO/EB（吖啶橙/溴化乙锭）和Hoechst 33258染色法研究了形态学凋亡。通过彗星试验检测DNA损伤。采用伤口愈合试验评估细胞迁移抑制情况。在荧光显微镜下研究了活性氧（ROS）水平和线粒体膜电位的变化。通过流式细胞术测定凋亡细胞和坏死细胞的百分比以及细胞周期阻滞情况。通过蛋白质免疫印迹分析研究了Bcl-2家族蛋白的表达。结果表明，这些配合物通过ROS介导的线粒体功能障碍途径诱导BEL-7402细胞凋亡，并伴有Bcl-2家族蛋白表达的调节。
