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目的检测PARP抑制剂维利帕尼(veliparib)与抗癌药物多柔比星(doxorubicin)联合应用对肝癌耐药细胞株BEL-7404增殖抑制的作用。方法以BEL-7404细胞为研究对象,常规培养传代,经多柔比星和(或)维利帕尼处理24h后,通过四甲基偶氮唑盐(MTT)比色法观察细胞的增殖率变化,采用流式细胞术Annexin V-FITC/PI双染法分析细胞凋亡水平,通过单细胞凝胶电泳实验评价DNA损伤程度,通过聚丙烯酰胺凝胶电泳(Western blotting)法检测细胞线粒体和胞浆中细胞色素c水平变化。结果多柔比星和维利帕尼联合应用组的细胞增殖率明显低于对照组和多柔比星单独处理组(P<0.01),细胞凋亡率则显著高于对照组和多柔比星单独处理组(P<0.05);同时,多柔比星和维利帕尼联合应用组细胞DNA损伤水平较对照组和多柔比星单独处理组明显加重(P<0.01),而细胞色素c在胞浆中的水平则显著高于对照组和多柔比星单独处理组(P<0.01)。结论维利帕尼与多柔比星联合作用可以抑制BEL-7404细胞的增殖,并诱导细胞DNA损伤和凋亡,其机制与胞浆中细胞色素c的表达升高有关。
Objective To investigate the inhibitory effect of a combination of PARP inhibitor veliparib and doxorubicin on the proliferation of multidrug resistance hepatocellular carcinoma cell line BEL-7404. Methods BEL-7404 cells were cultured in routine culture and passaged. After treated with doxorubicin and / or viliplatin for 24 h, the cell proliferation rate was observed by MTT assay. The cell apoptosis was analyzed by flow cytometry Annexin V-FITC / PI double staining. The degree of DNA damage was assessed by single cell gel electrophoresis. The mitochondrial and mitochondrial DNA levels were determined by polyacrylamide gel electrophoresis Changes of cytoplasm cytochrome c levels. Results Compared with the control group and doxorubicin alone group, the cell proliferation rates of doxorubicin and viliplatin combination group were significantly lower than those of the control group and the doxorubicin group (P <0.01), and the apoptosis rate was significantly higher than that of the control group and doxorubicin (P <0.05). At the same time, the DNA damage of the combination group of doxorubicin and vinpirin significantly increased compared with the control group and the doxorubicin alone group (P <0.01), while the cytochrome c in the cytoplasm was significantly higher than the control group and doxorubicin alone (P <0.01). Conclusion The combination of Velipan and doxorubicin can inhibit the proliferation of BEL-7404 cells and induce DNA damage and apoptosis. The mechanism is related to the increase of cytochrome c expression in cytoplasm.