钌配合物[Ru(MeIm)_4(bpy)]^(2+)的合成及对人肝癌细胞株增殖的影响

Synthesis of ruthenium complex [Ru(MeIm)_4(bpy)]^(2+) and its effect on proliferation in human liver cancer cell lines

目的合成钌配合物[Ru(MeIm)4(bpy)]2+并初步探讨其体外抗肝癌活性。方法利用元素分析、电喷雾质谱(ESI-MS)、核磁共振(NMR)等对目标化合物进行表征并通过溴化乙啶(EB)竞争结合实验检测其与DNA的结合能力;采用MTT法检测其对3株人肝癌细胞株HepG2、HCC-LM3和MHCC-97L细胞增殖的影响,用Hoechst33342染色法观察细胞凋亡的形态学改变,以流式细胞仪观察药物作用后细胞的凋亡率和细胞周期的变化。结果合成并表征了金属钌配合物[Ru(MeIm)4(bpy)]2+,同时竞争结合实验表明其能取代EB结合DNA。该化合物可抑制HepG2、HCC-LM3和MHCC-97L的增殖,呈浓度和时间依赖性。该化合物对HepG2细胞的增殖抑制作用最为明显,且浓度依赖性地诱导HepG2细胞凋亡,并随着作用时间的延长Sub-G1凋亡峰越增加。31.25～125μg/ml钌配合物阻滞细胞于G0/G1期,而浓度达250μg/ml时则阻滞细胞于G2/M期。结论合成的目标产物钌配合物[Ru(MeIm)4(bpy)]2+在体外可抑制人肝癌HepG2细胞的生长并能诱导其发生凋亡。

Objective To synthesize the ruthenium complex [Ru（MeIm）4（bpy）]^2＋ and study its in vitro activity against liver cancer.Methods The characteristics of [Ru（MeIm）4（bpy）] 2＋ were detected by elemental analysis,electrospray mass spectrometry（ESI-MS） and nuclear magnetic resonance（NMR）,its binding affinity for DNA was determined by competitive binding experiments,and its effect of proliferation of 3 human liver cancer cell lines（HepG2,HCC-LM3 and MHCC-97L） was assayed by MTT assay.Morphological changes of apoptotic human liver cancer cells were observed with Hoechst33342 staining.Changes in cell apoptosis rate and cell cycle after exposure to drugs were detected by flow cytometry.Results [Ru（MeIm）4（bpy）]^2＋ was synthesized and characterized in the present study.Competitive binding experiments showed that [Ru（MeIm）4（bpy）]^2＋ could bind with DNA instead of EB and inhibit the proliferation of HepG2,HCC-LM3 and MHCC-97L cells,especially of HepG2 cells,in a concentration-and time-dependent manner.[Ru（MeIm）4（bpy）]^2＋ could induce the apoptosis of HepG2 cells in a concentration-dependent manner and increase the apoptosis peak in Sub-G1 cells with the prolonged exposure to drugs.[Ru（MeIm）4（bpy）]^2＋ blocked the cell cycle at G0/G1 phase when its concentration was 31.25 to 125 μg/ml and arrested the cell cycle at G2/M phase when its concentration was 250 μg/ml.Conclusion Synthesized [Ru（MeIm）4（bpy）]^2＋ can inhibit the proliferation and induce apoptosis in HepG2 cells.