吡啶锰配合物诱导肿瘤细胞死亡的作用及对线粒体功能的影响

研究新合成的小分子吡啶锰配合物Adpa-Mn（III）（（Adpa）Mn（μ2-O）2Mn（Adpa）]PF6.8H2O（Adpa=bis（2-pyridylmethyl）amino-2-propionic acid））的抗肿瘤作用,初步探索其抗肿瘤的机制。MTT分析Adpa-Mn（III）对细胞活性的影响;活细胞工作站观察GFP荧光标记组蛋白HeLa细胞的细胞核形态,MDC染色以及GFP-LC3质粒转染,探讨细胞死亡的方式;JC-1染色检测线粒体膜电位;Fluo-3-AM和DCFH-DA荧光探针分别检测细胞中Ca^2＋和ROS的含量。结果发现,Adpa-Mn（III）剂量依赖性地抑制细胞活性;给药后细胞核出现固缩、片段化;自噬小泡增多,GFP-LC3荧光强度增强;线粒体膜电位下降;细胞内Ca^2＋发生超载,ROS含量升高。由此,Adpa-Mn（III）可抑制肿瘤细胞活性,其机制与引起线粒体膜电位下降、增加ROS生成及诱导细胞的死亡有关,同时胞内Ca^2＋超载也参与了该作用。这些数据显示,Adpa-Mn（III）具有成为抗肿瘤先导金属配合物的潜在可能性。

The Activity of Inducing Cancer Cell Death and Affecting Mitochondrial Function of a Novel Manganese-pyridine Compound

In this study,anticancer activity of the novel manganese-pyridine compound Adpa-Mn（III）（（Adpa）Mn（μ2-O）2Mn（Adpa）]PF6·8H2O（Adpa=bis（2-pyridylmethyl） amino-2-propionic acid）） and its possible mechanism were investigated.Four human cancer cell lines including HepG-2,HeLa,A549 and U251 cells were treated by manganese-pyridine derivative Adpa-Mn（III）.Cancer cell proliferation were detected by MTT assay.To observe cell apoptosis,the morphological and nuclei changes in H2B-GFP-labled HeLa cells were observed by a live cell system（LCS）.Autophagic cell death was studied with acidic vesicular organelles observation following monodansylcadervarine（MDC） labeling and autophagy-related proteins GFP-LC3 plasmid transfection.Mitochondrial membrane potential was observed by JC-1 staining;Intracellular free Ca^2＋ content was detected with Fluo-3 staining;Formation of ROS were detected by DCFH-DA staining.Our data show that Adpa-Mn（III） exhibited significant inhibition on cancer cell proliferation and exhibited dose-and time-dependent effect on U251 proliferation.Adpa-Mn（III） induced apoptosis indicated by chromatin condensation.Treartment of Adpa-Mn（III） enhanced fluorescence intensity of monodansylcadervarine（MDC） and GFP-LC3.Moreover,Adpa-Mn（III） induced mitochondrial membrane potential decreased,elevated ROS,and overloaded intracellular Ca^2＋.These results suggest that Adpa-Mn（III） exerts significant anticancer activity.Adpa-Mn（III） may induce apoptosis and autophagy of cancer cells.The possible mechanism underlying its anticancer effect was related to ROS-induced mitochondrial dysfunction.In summary,the current study suggest that Adpa-Mn（III） could be exploited as a potential lead compound as a novel anticancer metal-drug.