抗甲状腺药物致Graves病患者粒细胞缺乏高表达基因的筛选与克隆研究  

Screening and Cloning of Highly Expressed Genes in Patients of Graves' Disease with Agranulocytosis Result from Antithyroid Drugs

作　　者：钟警[1] 杨靖[1] 周斌[1] 刘江华[1] 曹仁贤[1] 文格波[1] 

机构地区：[1]南华大学第一附属医院临床医学研究所,湖南省衡阳市421001

出　　处：《中国全科医学》2010年第26期2921-2925,共5页Chinese General Practice

基　　金：国家自然科学基金(30840052);湖南省教育厅项目(07C627);湖南省卫生厅项目(C2006-025)

摘　　要：目的筛选可能对抗甲状腺药物(ATD)致Graves病(Graves disease,GD)患者粒细胞缺乏起决定作用的相关基因。方法采用淋巴细胞分离液分离GD服用ATD导致粒细胞缺乏患者与粒细胞正常患者的单个核细胞,加入EB病毒进行细胞培养,应用抑制性消减杂交(SSH)的方法构建GD服用ATD导致粒细胞缺乏患者与粒细胞正常患者之间差异表达的cDNA消减文库,克隆、鉴定ATD介导GD患者粒细胞缺乏特异性高表达的基因片段,以生物信息学方法进行初步分析。结果成功构建了人ATD介导GD患者粒细胞缺乏的cDNA消减文库;初步筛选到34条EST,其中21条为新EST,它们可能代表着在ATD介导GD患者粒细胞缺乏中特异性高表达并对ATD介导GD患者粒细胞缺乏的发生有促进作用的新基因。结论高效消减文库的成功建立,对进一步筛选、克隆GD差异表达的未知新基因和已知基因的新生物学功能研究具有重要意义。Objective To screen the highly expressed genes that may play a focal role in GD patients with agranulocytosis that results from anti-thyroid gland drugs.Methods Lymphocyte Separation Medium was used to separate mononuclear cells from patients of GD with agranulocytosis and normal patients.EB virus was used to culture the cells,and SSH approach was applied to construct cDNA subtractive library of differential expression between the patients of GD with agranulocytosis and normal patients.The highly expressed genes were cloned and sequenced.The sequencing results were compared in bioinformatics approach.Results A highly efficient subtractive cDNA library was established.34 ESTs were obtained,of which 21 clones were found to be novel ESTs as clues were associated with them by bioinformatic analysis.Conclusion The successful construction of subtractive cDNA library was significant for the intensive screening,cloning the Graves＇ disease＇s specific unknown new genes and studying the new biological functions of known genes.

关 键 词：抗甲状腺药 格雷夫斯病 粒细胞缺乏 抑制性消减杂交技术 基因 

分 类 号：R736.1[医药卫生—肿瘤]