脱氧雪腐镰刀菌烯醇致小鼠畸形骨骼组织差异表达基因及相关通路分析  

Differential gene expression and related pathways of skeletal malformations induced by deoxynivalenol in mice

作　　者：李岩[1] 赵英会[1] 庄东明[1] 李晓霞[1] 于爱莲[1] 

机构地区：[1]泰山医学院基础医学院,山东泰安271000

出　　处：《癌变．畸变．突变》2015年第1期1-5,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：山东省科技攻关项目(2007GG30002016);国家传染病重大专项合作项目(2009ZX10004-216);山东省自然科学基金(ZR2013HL060)

摘　　要：目的：DON致小鼠骨骼畸形的分子基础。方法：取孕期小鼠30只,随机分成3个不同浓度DON染毒组、溶剂对照组和空白对照组,每组6只,将DON染毒组孕鼠于孕期第7~10天连续腹腔注射DON,染毒后第18天麻醉剖腹取出胎鼠骨骼组织,提取椎骨骨骼组织总RNA,通过反转录法获取畸形骨骼组织和正常对照组骨骼组织的c DNA,并对其分别进行荧光标记,采用全基因组芯片技术对小鼠胚胎畸形骨骼组织与正常对照组织的差异基因表达谱进行分析,采用基因本体学分析软件对差异表达基因进行功能分析,DAVID database数据库筛选差异表达基因涉及的信号通路,并利用荧光定量PCR（q PCR）技术对基因芯片结果进行验证。结果：在DON染毒组小鼠全基因组芯片上筛选出差异基因282个,其中下调148个,上调134个;在差异表达基因分析的基础上,发现差异表达基因涉及到的通路有153个。q PCR结果表明,上调基因Fbn1 Co19a2 Papss2 Pax1F和下调基因Runx2 Pthlh等6个差异表达基因的相对定量结果与基因芯片检验结果表达趋势一致。结论：应用基因表达谱芯片初步筛选出DON所致胎鼠畸形骨骼与正常胎鼠骨骼组织中的差异表达基因,并通过通路分析发现,上述差异表达基因涉及多条信号传导通路。OBJECTIVE：To screen the differential gene expression and related pathways of fetal skeletal malformations induced by deoxynivalenol（DON） in mice,and to explore the mechanism of deformities induced by DON at the molecular level.METHODS：30 pregnant mice were randomly divided into 3 experimental groups and 2 control groups,6 in each group. DON injection was performed from days 7 to 10 of pregnancy by intraperitoneal injection into pregnant mice. At GD 18,all mice were killed under isoflurane anesthesia,and vertebral bone tissue of fetuses were collected. Total RNA of vertebral bone tissues was extracted and cDNA was obtained by RT-PCR. Using whole genome microarray technology,the gene expression profiles and the pathways of the rat vertebral bone tissue were studied. Analysis of the function of differentially expressed genes was performed by using software of gene ontology. Screening signaling pathways of differentially expressed genes was done by DAVID database. The results were verified by fluorescence quantitative PCR（qPCR） technology. RESULTS：Microarray analysis showed that 282 genes,including 148 down-regulated and 134 up-regulated genes,were abnormally expressed in fetal vertebral bones after maternal DON exposure. 153 pathways were related to the differentially expressed genes. The relative quantitative results ofqPCR were consistent w ith t he r esults o f g ene c hip t est i nFbn1,Co19a2,Papss2,Pax1,Runx2 a ndPthlh g enes.CONCLUSION： There were differential gene expressions in deformed fetal skeleton between deoxynivalenol-treated rats and normal rats, involving multiple signaling pathways. The differentially expressed genes and signaling pathways may be related to the molecular mechanisms of deformities induced by DON.

关 键 词：脱氧雪腐镰刀菌烯醇 骨骼畸形 基因芯片 差异表达基因 传导通路 

分 类 号：Q344.13[生物学—遗传学]