帕米膦酸二钠抑制人肝癌细胞增殖及诱导凋亡机制研究

发布时间：2018-08-09 14:08

【摘要】：目的肝癌是常见的恶性肿瘤之一,寻找其低毒有效的化疗药物具有重要的临床价值。本研究旨在探讨一种临床常用的骨吸收抑制剂帕米膦酸二钠对人肝癌HepG2及Bel-7402细胞体外增殖和凋亡的影响及其机制。方法采用MTS法检测不同浓度的帕米膦酸二钠对HepG2和Bel-7402细胞体外增殖的影响;以TUNEL末端标记法和Annexin-Ⅴ-FITC双标记流式法检测帕米膦酸二钠对细胞凋亡的影响采用蛋白质印迹法检测帕米膦酸二钠对凋亡相关蛋白:聚腺苷二磷酸-核糖聚合酶(poly ADP-ribose polymerase,PARP)、Caspase-9、Caspase-3、active-Caspase-3、Bax及Bcl-2表达的影响。结果 MTS检测显示,帕米膦酸二钠对HepG2及Bel-7402细胞均具有明显抑制作用,并呈剂量依赖性,药物的半数抑制浓度(50%inhibitory concentration,IC_(50))分别为46.12和51.35μmol/L。流式细胞术检测发现,帕米膦酸二钠可诱导肝癌细胞剂量依赖性凋亡,20、40和60μmol/L作用HepG2细胞24h凋亡率分别为(25.70±6.00)%、(43.53±4.06)%和(74.73±2.90)%,与对照组的(3.43±1.07)%相比差异有统计学意义,F=175.5,P0.001;20、40和60μmol/L作用Bel-7402细胞24h凋亡率分别为(25.50±3.00)%、(48.30±1.02)%和(68.97±4.90)%,与对照组(3.33±0.05)%相比差异有统计学意义,F=81.97,P0.001。蛋白质印迹法检测结果显示,帕米膦酸二钠可诱导细胞内凋亡相关蛋白Caspase-9、Caspase-3及PARP的活化,促凋亡蛋白Bax的上调,抑凋亡蛋白Bcl-2的下调。结论帕米膦酸二钠可以明显抑制人肝癌HepG2及Bel-7402细胞的体外增殖,其机制可能与线粒体凋亡途径的激活而诱导癌细胞发生凋亡相关,帕米膦酸二钠可能成为潜在的肝癌治疗药物。
[Abstract]:Objective Hepatocellular carcinoma is one of the most common malignant tumors, and it has important clinical value to search for its low toxicity and effective chemotherapeutic drugs. The aim of this study was to investigate the effects of a common bone resorption inhibitor, Pamidronate disodium, on the proliferation and apoptosis of human hepatocellular carcinoma (HepG2) and Bel-7402 cells in vitro and its mechanism. Methods the effects of disodium Pamidronate at different concentrations on the proliferation of HepG2 and Bel-7402 cells in vitro were detected by MTS assay. Effect of disodium Pamidronate on cell apoptosis by TUNEL end labeling and Annexin- 鈪，

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