目的：构建组蛋白去乙酰化酶（histone deacetylase 1, HDAC1）过表达胶质瘤U87细胞系，探讨HDAC1过表达对胶质瘤细胞化疗药耐药性的影响。方法：分别用HDAC1重组过表达慢病毒载体pCDH-CMV-MCS-HDAC1-EF1-Puro和阴性空载体病毒pCDH-CMV-MCS-EF1-Puro 转染U87MG细胞，通过嘌呤梯度浓度筛选出HDAC1稳定过表达细胞系U87MG-HDAC1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，MTT法检测存活率，Hoechst/PI双染法检测细胞凋亡，Western Blotting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-HDAC1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\[01μg/ml VM-26：（9557±0.45）% vs （68.8±1.49）%，P<0.01\]，\[0.08 μg/ml DDP：（99.20±7.4）% vs（72.48±2.03）%，P<0.01\]；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

Objective:To explore the effect of histone deacetylase 1 (HDAC1) over-expression on drug resistance of glioma cells. Methods: U87MG cells were infected with a recombinant lentiviral vector pCDH-CMV-MCS-HDAC1-EF1-Puro/over-expressing HDAC1 and a control lentiviral pCDH-CMV-MCS-EF1-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 successfuly constructed. Compared with U87GM-Control group, the expression of U87MG-HDAC1 protein was significantly increased 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 cells to chemotherapeutic drugs, which was may associated with the expressions of Caspase-3 and Bcl-2/Bax.

国家自然科学基金青年科学基金资助项目（No. 30701030）；天津市应用基础及前沿技术研究计划资助（No. 11JCYBJ14900）