靶向上调ID4基因表达药物的生物信息学预测和初步验证  

Experimental identification of drugs with function of targeted up-regulating ID4 expression： bioinformatics-based prediction and preliminary validation

作　　者：杨波[1] 卢学春[1] 刘丽宏 朱宏丽[1] 迟小华 姚善谦[1] 楼方定[2] 于力[2] 

机构地区：[1]解放军总医院老年血液科,北京100853 [2]解放军总医院血液科,北京100853 [3]解放军第二六二医院药剂科

出　　处：《中华医学杂志》2009年第24期1714-1716,共3页National Medical Journal of China

基　　金：国家自然科学基金（30772597、30873086）

摘　　要：目的筛选具有靶向上调ID4基因表达的药物。方法钓取ID4基因5’侧翼区3000bp非编码DNA序列和mRNA序列中的开放阅读框（ORF）；利用启动子分析软件在人类转录因子数据库中搜索可能存在的顺式结构；利用基因表达数据库分析影响ID4基因表达的相关药物，并做药物调控人类基因表达谱的相似性分析。将筛选出的活性药物与急性淋巴细胞白血病细胞系MOLT4共培养，RT—PCR方法检测IIM基因在药物作用前后的表达情况。结果ID4基因具有Ⅱ型启动子，在5’非翻译区的-45bp处有一个典型的TATA盒。在长度为1300bp的ID4启动子区，存在多个顺式结构，其中转录因子Sp1、c-Myb、环磷腺苷、糖皮质激素受体和雌激素受体可能具有正向调控作用；Wilms瘤基因（WT1）和早期生长反应基因2（EGR2）可能具有负向调控作用。环磷腺苷与ID4基因对人类基因表达谱的调控具有相似性。浓度为0．1mmol/L的二丁酰环磷腺苷具有诱导MOLT4细胞表达ID4基因的作用。结论启动子调控序列预测、基因表达谱相似性分析和文献检索相结合的生物信息学分析法有助于寻找调控目的基因表达的药物。二丁酰环磷腺苷具有诱导ID4基因在白血病细胞中表达的作用。Objective To screen new candidate molecular-targeted anti-leukemia compounds with potential functions of targeted up-regulating ID4 gene expression. Methods Promoter region of ID4 gene including the upstream - 3000 bp sequence of transcriptional start site and message RNA sequence were fished out. Online promoter analysis tools of TESS and Genomax were used to search possible sequence of transcriptional start site and message RNA sequence were fished out. Online promoter analysis tools of TESS and Genomax were used to search possible cis-acting structure from human transcription factor database. The activity of related drugs with potential effects upon ID4 gene expression was analyzed using SAGE database. GEO database was applied to search the gene expression profiling regulated by ID4 gene. Finally, similar analysis between gene expression profiling by ID4 and genome-wide profiling regulated by 163 known drugs or active compounds was manipulated to screen the drugs and candidate compounds with similar gene expression profiling with ID4 gene. MOLT4 cell line was treated with the above candidate active compounds to investigate the ID4 gene expression by RT-PCR assay. Results ID4 gene had a type Ⅱ promoter with a typical TATA box in upstream -45 bp of transcription start site. The 1300 bp-length promoter of ID4 gene contained a few cis- acting structures classified into two function types, i. e. positive regulatory type, including transcription factors Spl and c-Myb, cAMP, glucocorticoid receptor （GR） and estrogen receptor （ER） , and negative regulatory type, including Wilms tumor-1 （WT1） and early growth response-2 （EGR2）. The similarity of gene expression profiling was identified between cAMP and ID4 gene. ID4 gene expression was induced in MOLT4 cell line after treatment with calcium dibutyryladenosine cyclophosphate at the concentration of 0. 1 mmol/L. Conclusion The comprehensive bioinformatic analysis, based upon the combination of regulatory sequence prediction of promoter, similarity analy

关 键 词：白血病 计算生物学 基因 ID4 

分 类 号：R686[医药卫生—骨科学]