【摘要】 目的:通过筛选小鼠下颌下腺发育起关键作用的基因,并预测其基因功能,为唾液腺组织工程奠定实验基础。方法:选取小鼠胚胎第18.5天、19.5天、出生后当天、出生后3天等4个不同时期的下颌下腺,进行基因芯片检测,BeadStation 500 System扫描仪(illumina,Inc.)对芯片扫描,然后采用BeadStudio Gene Expression Module图像分析软件(illumina,Inc.)对芯片灰度扫描图进行分析,构建小鼠下颌下腺的全基因表达谱。应用生物信息学STC法分析基因表达趋势,建立发育时间与基因表达相关的调控网络图,从网络中筛选关键基因,预测其基因功能,并对关键基因Skp2进行实时荧光定量PCR验证。结果:应用STC生物学方法,得到差异基因的8种显著性表达趋势,其中有4种与表型相关。构建网络图筛选关键基因,得到Ddx1、Cenpl、Fanci、Skp2、Rbm4、Dak。Skp2基因检测数据与实时荧光定量PCR验证结果一致。其中Skp2与胚胎期细胞的分化、增殖密切相关。结论:6个关键基因在小鼠下颌下腺发育过程中起重要作用。更多还原

【Abstract】 PURPOSE:To screen the Key genes during salivary gland development in mice and predict the functions,so as to provide the promising strategy for the salivary gland tissue engineering.METHODS:Four submandibular gland samples were chosen at selected times(E18.5,E19.5,P0,P3) during development of the murine.By mRNA hybridization,BeadStation 500 System scanner and illumine BeadStudio application,mRNA expression profiles were established.Then,using series time cluster(STC)and multiple comparison tests,a regulation network was constructed according to gene expression in the course of submandibular gland development.The key genes from the network were screened,and the functions were predicted.Microarray data was validated using real-time PCR.RESULTS:The results showed that using STC,eight significant expression strands were established among the different genes,four of which were closely correlated with phenotype.The network was established and the key genes were Ddx1,Cenpl,Fanci,Skp2,Rbm4,Dak.Microarrays and real-time PCR data was in accordance.Skp2 was much relevant to cell differentiation and proliferation during embryo stage in mice.CONCLUSION:Ddx11,Cenpl,Fanci,Skp2,Rbm4,Dak play an important role in submandibular gland development in mice,and the study can also be used for further research in the salivary gland tissue engineering.更多还原