中文摘要：

目的：构建组蛋白去乙酰化酶（histone deacetylase 1，HDACl）过表达胶质瘤U87细胞系，探讨HDACl过表达对胶质瘤细胞化疗药耐药性的影响。方法：分别用HDACl重组过表达慢病毒载体pCDH-CMV-MCS-HDACl-EFl-Puro和阴性空载体病毒pCDH-CMV-MCS-EFl-Puro转染U87MG细胞，通过嘌呤梯度浓度筛选出IqDACl稳定过表达细胞系U87MG-HDACl和空载体对照细胞系U87MG-Control，经Western Blotting鉴定，用不同浓度替尼泊苷（VM-26）和顺铂（cisplatin，DDP）对两种细胞进行处理，MTY法检测存活率，Hoechst／PI双染法检测细胞凋亡，WesternBlotting检测Bcl-2、Bax及Caspase-3蛋白表达。结果：成功构建U87MG-HDAC1稳定过表达细胞系。与U87MG-Control组相比，U87MG-HDAC1组中U87MG-HDACl蛋白的表达显著升高[（1．148±0．024）vs（0．580±0．003），P〈0．01]；药物处理后，U87MG-HDAC1细胞存活率显著升高[0．1μg／ml VM-26：（95．57±0．45）％vs（68．8±1．49）％，P〈0．01l，[0．08μg／ml DDP：（99．20±7．4）％似（72．48±2．03）％，P〈0．01]；凋亡细胞个数比例明显下降（均P〈0．05），Caspase-3蛋白表达量显著降低（P〈0．01），Bcl-2／Bax蛋白表达量比值明显升高（P〈0．01）。结论：胶质瘤U87MG细胞中HDAC1过表达明显增强细胞对化疗药耐药性与Caspase-3和Bcl-2／Bax表达有关。

英文摘要：

Objective：To explore the effect of histone deacetylase 1 （HDAC1） over-expression on drug resistance of glioma cells. Methods： U87MG ceils were infected with a recombinant lentiviral vector pCDH-CMV-MCS-HDAC1-EF1- Puro/over-expressing HDAC1 and a control lentiviral pCDH-CMV-MCS-EFI-Puro, respectively. Cell line U87MG- HDAC1 which stably over expressing HDAC1, and control cell line U87MG-Control were selected with purine gradient concentration and identified by Western blotting. After processed with VM-26 and DDP at different concentrations assessed by MTT of the cells was evaluated by Hoechst/PI double staining assay, and the expressions of Bcl-2,Bax and Caspase-3 were examined by Western blotting. Results： Cell line U87MG-HDAC1 that stably over expressing HDAC1 was successfu- ly constructed. Compared with U87GM-Control group, the expression of U87MG-HDAC1 protein was significantly in- creased in U87MG-HDAC1 group （ [ 1. 148± 0.024 ] vs [ 0.580 ± 0. 003 ], P 〈 0.01 ）. After drug treatment the survival rate of U87MG-HDAC1 cell was remarkably higher （0.1 μg/ml VM-26 ： [ 95.57 ± 0.45 ] % vs [ 68.8 ± 1.49 ] %, P 〈 0. 01 ） ; （0.08 μg/ml DDP： [ 99.20± 7.4 ] % vs [ 72.48 ± 2.03 ] %, P 〈 0.01 ） ; Number of apoptotic cells decreased （P 〈 0.05） ; the expression of Caspase-3 protein decreased significantly （P 〈 0.01 ） and the expression ratio of Bcl-2/ Bax proteins was significantly enhanced （P 〈 0.01 ）. Conclusion： The over-expression of HDAC1 in glima cell U87MG remarkably enhance of resistance of the ceils to chemotherapeutic drugs, which was mav associated with the exnressions of Caspase-3 and Bcl-2/Bax.