【摘要】 目的观察聚腺苷二磷酸核糖聚合酶(PARP)-1抑制剂AG-014699和AZD2281对人肝癌细胞株Hep G2细胞抑制作用和凋亡,初步探讨PARP-1抑制剂诱导Hep G2细胞凋亡的机制,为肝癌提供一种新的治疗靶点。方法 MTT实验观察不同浓度的AG-014699和AZD2281对Hep G2细胞增殖的影响,用流式细胞术检测Hep G2细胞凋亡率;Western Blot法检测casepase3和casepase8蛋白表达水平。组间比较采用t检验。结果 AG014699和AZD2281均有抑制Hep G2细胞增殖的作用,且具有时间和浓度依赖性,但Hep G2细胞对两种PARP-1抑制剂的敏感性不同,用MTT法检测48 h AG-014699和AZD2281的IC50分别约为20、400μmol/L。因AZD2281不敏感,未做流式细胞术和Western Blot法检测细胞凋亡。用10、30、50μmol/L的AG-014699能诱导Hep G2细胞凋亡,48 h时凋亡率最高达(31.00±2.13)%,明显高于对照组(0.900±0.013)%,二者差异有统计学意义(P<0.01)。30、50μmol/L的AG-014699作用Hep G2细胞48 h后的caspase3和caspase8蛋白水平相对于正常对照组显著增加。结论 PARP-1抑制剂AG-014699和AZD2281均能抑制Hep G2细胞增殖,但敏感性不同,AG-014699可诱导Hep G2细胞凋亡,通过上调caspase3和caspase8蛋白水平来诱导细胞凋亡。更多还原

【Abstract】 Objective To observe the inhibitory and pro- apoptotic effects of two poly( ADP- ribose) polymerase( PARP- 1) inhibitors,AG- 014699 and AZD2281,on human hepatoma Hep G2 cells and preliminarily explore the mechanism by which AG- 014699 induces Hep G2 cell apoptosis,and to provide a new therapeutic target for hepatoma. Methods The effects of different concentrations of AG-014699 and AZD2281 on Hep G2 cell proliferation were determined by MTT assay. The cell apoptosis rate was measured by flow cytometry.The expression levels of caspase- 3 and caspase- 8 were measured by Western Blot. Inter- group comparison was made by t test. Results Both AG- 014699 and AZD2281 suppressed Hep G2 cell proliferation in a time- and dose- dependent manner. However,the sensitivity of Hep G2 cells to the two PARP- 1 inhibitors was different. The half- maximal inhibitory concentrations of AG- 014699 and AZD2281 at48 h determined by MTT assay were about 20 μmol / L and 400 μmol / L,respectively. Flow cytometry and Western blot were not used to evaluate the apoptosis of Hep G2 cells exposed to AZD2281 to which these cells were not sensitive. Hep G2 cell apoptosis could be induced by10,30,and 50 μmol / L AG- 014699,and the highest apoptosis rate at 48 h was significantly higher than that of the control group( 31. 00% ± 2. 13% vs 0. 9% ± 0. 013%,P < 0. 01). Compared with those in the control group,the protein levels of caspase- 3 and caspase- 8 in Hep G2 cells after 48- h exposure to 30,and 50 μmol / L AG- 014699 increased. Conclusion The two PARP- 1 inhibitors AG- 014699 and AZD2281 can inhibit the proliferation of Hep G2 cells,which showed different sensitivities to the two inhibitors. AG-014699 can induce Hep G2 cell apoptosis by up- regulating the protein expression of caspase- 3 and caspase- 8.更多还原