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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。
OBJECTIVE: To lay the foundation for the salivary gland tissue engineering by screening the genes that play a key role in the development of mouse submandibular gland and predicting their gene function. METHODS: The submandibular gland of four different periods including embryonic day 18.5, 19.5, postnatal day 3, and postnatal day 3 were selected for microarray detection. Microarray scanning was performed with the BeadStation 500 System scanner (illumina, Inc.) The gray scale scan of the chip was analyzed by using the BeadStudio Gene Expression Module image analysis software (Illumina, Inc.) to construct the whole gene expression profile of the mouse submandibular gland. The bioinformatics STC method was used to analyze the trend of gene expression and to establish the regulatory network map related to gene expression during developmental time. The key genes were screened from the network to predict the gene function, and the key gene Skp2 was verified by real-time fluorescence quantitative PCR. Results: Eight kinds of significant expression trends of the differentially expressed genes were obtained by using STC biological methods, of which four were related to the phenotype. Construction of network map screening of key genes, get Ddx1, Cenpl, Fanci, Skp2, Rbm4, Dak. Skp2 gene detection data and real-time fluorescence quantitative PCR validation results. Which Skp2 and embryonic cell differentiation, proliferation is closely related. Conclusion: Six key genes play an important role in the development of mouse submandibular gland.