表达谱基因芯片筛选巨指脂肪组织差异表达基因  

Identification of differential expression genes related to fatty tissue of macrodactyly by gene expression profiles chip

作　　者：李文军[1] 常会波[2] 田光磊[1] 陈山林[1] 田文[1] 赵俊会[1] 杨勇[1] 

机构地区：[1]北京积水潭医院手外科,100035 [2]首都儿科研究所生物化学研究室

出　　处：《中华手外科杂志》2015年第6期463-466,共4页Chinese Journal of Hand Surgery

基　　金：国家临床重点专科北京积水潭医院手外科专项基金资助项目[（2010）305号]

摘　　要：目的利用表达谱基因芯片初步研究巨指脂肪组织mRNA表达谱差异性，并筛选验证差异基因，为后续研究提供基础数据。方法自2012年4月至12月，我们共收集5例巨指症患儿正常和病变脂肪组织。提取组织RNA后，检测总RNA的质量和纯度，合格样品RNA反转录制备含荧光分子cv5标记的cDNA探针，与含有30968点cDNA的表达谱芯片杂交后扫描荧光强度，筛选出差异表达的基因并进行分析，选取5个基因利用实时定量PCR技术验证差异表达。结果使用mRNA表达谱芯片筛选出了1725个差异表达基因，其中上调基因801个，下调基因924个。实时定量PCR结果显示4个基因表达上调，1个基因表达下调，表达趋势与芯片结果一致。结论巨指症患儿中受累脂肪组织和正常脂肪组织基因表达存在差异，分析这些表达基因有助于我们探索巨指发病机制，为疾病治疗提供理论依据。Objective To detect and compare the gene expression profiles of normal adipose tissue and adipose tissue from maerodctyly fingers using eDNA microarray method and screen differential expression genes related to macrodactyly to provide basic data for future studies. Melhods Normal adipose tissue and adipose tissue from the affected fingers were collected from 5 patients with macrodaetyly who were treated between April and December of 2012. Total R_NA was extracted and purified for mRNA expression profile study. According to Trizol instructions for RNA extraction, the samples were analyzed with Agilent 2100 Bioanalyzer to detemfine the quantity and purity of RNA. RNA samples were used only if the ratio of A260 to A280 values fell within the range of 1.7 to 2.1, and the RNA integrity number was greater than 8.0. cDNA probes containing fluorescent molecular Cy5 markers were prepared from the RNA samples via reverse transcription and applied to a eDNA chip containing 30968 spots. The fluorescence intensity of chip hybridization was scarmed to screen and identify genes that were differentially expressed. The expression levels of 5 randomly chosen genes were verified by quantitative RT-PCR. Results A total of 1725 genes were differentially expressed in in the fatty tissue of macrodaetyly. Among them 801 were up-regulated and 924 down-regulated. The expression trend of the 5 genes chosen for further verification was consistent with the q-PCR results （4 up-regulated and 1 down-regulated）. Conclusion In macrodactyly patients, there are distinctly different gene expression profiles of affected adipose tissue when compared to the normal adipose tissue. Analyzing these genes may help clarify the pathogenesis of macrodactyly and provide basis for trealment of the disease.

关 键 词：脂肪组织 基因表达谱 巨指症 差异表达基因 

分 类 号：R735.7[医药卫生—肿瘤]