摘要
目的探讨先天性心脏病相关基因转录因子Pax-8的下游基因以及Pax-8基因下调引起心脏形态改变的机制。方法分别提取Pax-8基因敲除小鼠纯合子（Pax-8 KO^-/-）和杂合子（Pax-8 KO^4/-）的心脏总RNA，利用含31802个小鼠基因的基因芯片检测Pax-8 KO^-/-和Pax-8 KO^-/-小鼠的基因表达水平，找出差异表达的基因，并经荧光实时定量PCR技术初步筛选出转录因子Pax-8的下游基因。结果基因芯片检测发现，Pax-8 KO^-/-小鼠较Pax-8 KO^＋／-小鼠有25个基因表达下调，另有17个基因表达上调，差异基因涉及细胞周期及信号转导的调节因子、直接参与代谢的酶以及核转录因子等。定量RT-PCR证实，Pax-8 KO^-/-小鼠Fgf-6基因的表达水平较Pax-8 KO^＋/-小鼠及野生型（Pax-8^＋/＋）小鼠分别上调1.13倍和1．67倍，差异均较显著（P〈0.01）。结论Fgf-6基因是转录因子Pax-8的下游基因，可能在心脏的发育过程中发挥着重要作用。
Objective To investigate the downstream genes of the transcriptional factor-Pax-8 related to congenital heart disease. Methods The total RNA was extracted from the heart of Pax-8 KO^-/- and Pax-8 KO^＋/- mice. Mouse genome DNA microarray containing 31,802 mouse oligonucleotides probes was used to examine the differential expression pattern between the Pax-8 KO^-/- and the Pax-8 KO^＋/-. The candidate genes were confirmed by real time RT-PCR assay. Results Microarray results showed that compared to Pax-8 KO^＋/- mice, the expression level of 25 genes was down-regulated and that of 17 genes was up-regulated in Pax-8 KO ^-/- mice, those genes were related to metabolize enzyme, cell signal conduction and nuclear transcript factors. Real time RT-PCR results revealed that fibroblast growth factors-6 （Fgf-6） in the Pax-8 KO^-/- mice was 1.13 and 1.67 fold higher as that in Pax-8 KO^＋/- and Pax-8^＋/＋ mice （P 〈0.01）.Conclusion The Fgf-6 gene was one of the downstream genes of the Pax-8 and probably was involved in the heart development.
出处
《浙江医学》
CAS
2010年第11期1591-1593,共3页
Zhejiang Medical Journal
基金
国家自然科学基金（30571050）
浙江省温州市科技发展计划资助项目（Y2006A017）
（致谢：此课题的完成得到德国Peter Gruss及Ahmed Mansouri教授的支持和帮助,尤其是惠赠实验动物,特此表示感谢!）
