HDAC1过表达诱导胶质瘤细胞产生耐药性  

Drug resistance of glioma cells induced by over-expression of HDAC1

作　　者：郭谦[1] 曹蕾[1] 卓小煌 王彰昭 张欢[1] 强兆艳[1] 何景华[1] 

机构地区：[1]天津医科大学基础医学院药理学教研室,天津300070

出　　处：《中国肿瘤生物治疗杂志》2016年第1期67-72,共6页Chinese Journal of Cancer Biotherapy

基　　金：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)~~

摘　　要：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p CDH-CMV-MCS-HDAC1-EF1-Puro和阴性空载体病毒p CDH-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U87MGHDAC1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],[0.08μg/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 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.

关 键 词：组蛋白去乙酰化酶 胶质瘤 耐药性 

分 类 号：R739.41[医药卫生—肿瘤] R730.54[医药卫生—临床医学]