【摘要】：目的肝癌是常見的惡性腫瘤之一,尋找其低毒有效的化療藥物具有重要的臨床價值。本研究旨在探討一種臨床常用的骨吸收抑制劑帕米膦酸二鈉對人肝癌HepG2及Bel-7402細胞體外增殖和凋亡的影響及其機制。方法采用MTS法檢測不同濃度的帕米膦酸二鈉對HepG2和Bel-7402細胞體外增殖的影響;以TUNEL末端標(biāo)記法和Annexin-Ⅴ-FITC雙標(biāo)記流式法檢測帕米膦酸二鈉對細胞凋亡的影響采用蛋白質(zhì)印跡法檢測帕米膦酸二鈉對凋亡相關(guān)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表達的影響。結(jié)果 MTS檢測顯示,帕米膦酸二鈉對HepG2及Bel-7402細胞均具有明顯抑制作用,并呈劑量依賴性,藥物的半數(shù)抑制濃度(50%inhibitory concentration,IC_(50))分別為46.12和51.35μmol/L。流式細胞術(shù)檢測發(fā)現(xiàn),帕米膦酸二鈉可誘導(dǎo)肝癌細胞劑量依賴性凋亡,20、40和60μmol/L作用HepG2細胞24h凋亡率分別為(25.70±6.00)%、(43.53±4.06)%和(74.73±2.90)%,與對照組的(3.43±1.07)%相比差異有統(tǒng)計學(xué)意義,F=175.5,P0.001;20、40和60μmol/L作用Bel-7402細胞24h凋亡率分別為(25.50±3.00)%、(48.30±1.02)%和(68.97±4.90)%,與對照組(3.33±0.05)%相比差異有統(tǒng)計學(xué)意義,F=81.97,P0.001。蛋白質(zhì)印跡法檢測結(jié)果顯示,帕米膦酸二鈉可誘導(dǎo)細胞內(nèi)凋亡相關(guān)蛋白Caspase-9、Caspase-3及PARP的活化,促凋亡蛋白Bax的上調(diào),抑凋亡蛋白Bcl-2的下調(diào)。結(jié)論帕米膦酸二鈉可以明顯抑制人肝癌HepG2及Bel-7402細胞的體外增殖,其機制可能與線粒體凋亡途徑的激活而誘導(dǎo)癌細胞發(fā)生凋亡相關(guān),帕米膦酸二鈉可能成為潛在的肝癌治療藥物。
[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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