组蛋白去乙酰化酶1相互作用蛋白的筛选  

Screening of potential proteins interacting with histone deacetylase 1

作　　者：陈霞[1] 曾严[2] 张毅[2] 庞学利[3] 张杰峰[2] 

机构地区：[1]成都大学附属医院儿科,四川成都610081 [2]重庆市肿瘤研究所肝胆外科,重庆400030 [3]第三军医大学西南医院肿瘤科,重庆400038

出　　处：《现代肿瘤医学》2017年第4期516-520,共5页Journal of Modern Oncology

基　　金：国家自然科学基金面上项目(编号:81272497)

摘　　要：目的:筛选与组蛋白去乙酰化酶1(histone deacetylase 1,HDAC1)相互作用蛋白。方法:将带有Flag标签的HDAC1表达质粒分别转染食管癌细胞EC109和结肠癌细胞HCT116,采用免疫印迹检测外源基因的表达。然后采用Flag抗体进行亲和沉淀,收集蛋白样品进行质谱测定。结果:采用RT-PCR技术成功地构建了含Flag标签的HDAC1。转染后检测显示该表达质粒可高效表达于EC109和HCT116细胞中。亲和沉淀所获得的蛋白样品经质谱检测后,分别从EC109细胞和HCT116细胞中筛选出107和17个可能与HDAC1相互作用的蛋白质。结论:通过质谱筛选出可能与HDAC1相互作用的蛋白,为进一步研究HDAC1基因功能及其在肿瘤中的生物学意义奠定基础。Objective： To screen potential proteins interacting with histone deacetylase 1（ HDAC1）. Methods：Expression plasmid of Flag- tagged HDAC1 was constructed and transfected into esophageal cell line EC109 and colon cancer cell line HCT116. After transfection,cells were harvested and protein expression of HDAC1 was detected by Western blotting. Protein samples of immunoprecipitation by using Flag antibody were analyzed by mass spectrum.Results： Flag- tagged HDAC1 was successfully cloned into expression vector by RT- PCR. Forty- eight hours after transfection,Flag- tagged HDAC1 was efficiently expressed in both cell lines. Sufficient protein samples were obtained by immunoprecipitation with Flag antibody. After measured by mass spectrum and database retrieving,17 and107 potential proteins interacting with HDAC1 were obtained from EC109 and HCT116 cells,respectively. Conclusion： Potential proteins interacting with HDAC1 were successfully selected from two cancer cell lines,which contributed to further analysis of the gene functions of HDAC1 and its biological significance in cancer cells.

关 键 词：组蛋白去乙酰化酶1 食管癌细胞 结肠癌细胞 质谱 

分 类 号：R735.1[医药卫生—肿瘤] R735.35[医药卫生—临床医学]