经典WNT信号通路关键基因在人胎肺中的表达

发布时间：2018-05-20 11:52

本文选题：经典WNT信号通路 + 人胎肺　；参考：《福建师范大学》2012年硕士论文

【摘要】：已知经典WNT信号通路通过参与细胞的增殖、凋亡和分化过程,对小鼠肺器官发育起着重要的调控作用,但经典WNT信号通路在人胎肺发育时期的表达模式及调控功能目前未见相关报道。为此,本论文选取假腺肺(7W、12W、17W)和微管肺(21W)时期的人胎肺组织,通过Real-time PCR和原位杂交技术,检测经典WNT信号通路关键基因,包括WNT配体(WNT2、WNT7B),WNT受体(FZD4、FZD7、LRP5、LRP6),WNT信号通路调控器(DVL2、DVL3、GSK-3β, β-CATENIN、APC,AXIN2),WNT信号通路下游转录因子(TCF4、LEF1)以及WNT信号通路拮抗物(SOSTDC1)的表达模式,为阐明经典WNT信号通路调控人肺器官发育机制奠定基础。首先Real-time PCR结果显示,经典WNT信号通路关键基因在人胎肺发育4个时期(7W、12W、17W和21W)均有表达,其中WNT2.D VL2、GSK-3β,β-CATENIN,APC、TCF4及LEF1从7W到12W表达量逐渐下调,从12W到17W开始升高,但在21W又显著下调；LRP5、LRP6、DVL3、AXIN2和SOSTDC1从7W、12W到17W表达量逐渐升高,但到21W又显著下调；WNT7B和FZD7从7W到12W表达量升高,但从12W到21W又逐渐下降。其次原位杂交结果显示,除了经典WNT信号通路调控器DVL3、β-CATENIN, AXIN2及下游转录因子TCF4在胎肺近端气管及远端肺泡上皮和间充质中均有表达外,其余经典WNT信号通路调控器仅在胎月市气管分支及远端肺泡上皮表达。此外,我们利用小分子化合物CHIR-99021刺激15周的人胎肺组织以激活经典WNT信号通路,Western Blot、Real-time PCR及组织病理检测证实经典WNT信号通路被激活,并且过度激活的WNT信号致使人胎肺近远端肺部上皮细胞由矮柱状向高柱状的早期细胞状态退化。总之,我们的研究表明,经典WNT信号通路对人胎肺气管的分支形态及上皮细胞的分化均起着重要的调控作用,这将为研究经典WNT信号通路调控人胎肺发育的分子机制及相关肺部疾病的诊疗奠定基础。
[Abstract]:Classical WNT signaling pathway plays an important role in regulating the development of mouse lung organs by participating in cell proliferation, apoptosis and differentiation. However, the expression pattern and regulatory function of classical WNT signaling pathway during the development of human fetal lung have not been reported. In this paper, the human fetal lung tissues in the period of 7 W7 WN 12 WN 17 W) and 21 W microtubule lung were selected, and the key genes of classical WNT signaling pathway were detected by Real-time PCR and in situ hybridization. The expression pattern of WNT ligand WNT2 / WNT7BN / WNT receptor FZD4 / FZD7 / LRP5 / LRP6 / WNT signal pathway regulator DVL2DVL3 尾, 尾 -CATENINAPCAXIN2WNT signal pathway downstream transcription factor TCF4 / LEF1 and WNT signal pathway antagonist, SOSTDC1), is the basis for elucidating the regulation of human lung organ development by classical WNT signaling pathway. The results of Real-time PCR showed that the key genes of classical WNT signaling pathway were expressed at 7W ~ 12W ~ (17W) and 21W at four stages of human fetal lung development. The expression of GSK-3 尾, 尾 -CATENINAPC-TCF4 and LEF1 were down-regulated from 7W to 12W, and increased from 12W to 17W. However, at 21W, the expression of DVL3AXIN2 and SOSTDC1 increased gradually from 7W to 17W, but at 21W, the expression of WNT7B and FZD7 increased from 7W to 12W, but decreased gradually from 12W to 21W. The results of in situ hybridization showed that, in addition to the classical WNT signal pathway regulators DVL3, 尾 -CATENIN, AXIN2 and downstream transcription factor TCF4, they were expressed in the proximal and distal alveolar epithelium and mesenchyme of fetal lung. The other classical WNT signaling pathway regulators were only expressed in tracheal branches and distal alveolar epithelium. In addition, CHIR-99021 was used to stimulate human fetal lung tissue for 15 weeks in order to activate the classical WNT signaling pathway, Western blotte Real-time PCR and histopathological examination, which confirmed that the classical WNT signaling pathway was activated. And the overexpression of WNT signal degenerated the epithelial cells from short columnar to high columnar lung epithelial cells in the proximal and distal lung of human fetal lung. In conclusion, our study shows that the classical WNT signaling pathway plays an important role in regulating the branching morphology and epithelial cell differentiation of human fetal pulmonary duct. This will lay a foundation for the study of the molecular mechanism of classical WNT signaling pathway regulating human fetal lung development and the diagnosis and treatment of lung diseases.
【学位授予单位】：福建师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R321

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