本文選題：經(jīng)典WNT信號通路 + 人胎肺　； 參考：《福建師范大學(xué)》2012年碩士論文

【摘要】：已知經(jīng)典WNT信號通路通過參與細(xì)胞的增殖、凋亡和分化過程,對小鼠肺器官發(fā)育起著重要的調(diào)控作用,但經(jīng)典WNT信號通路在人胎肺發(fā)育時(shí)期的表達(dá)模式及調(diào)控功能目前未見相關(guān)報(bào)道。為此,本論文選取假腺肺(7W、12W、17W)和微管肺(21W)時(shí)期的人胎肺組織,通過Real-time PCR和原位雜交技術(shù),檢測經(jīng)典WNT信號通路關(guān)鍵基因,包括WNT配體(WNT2、WNT7B),WNT受體(FZD4、FZD7、LRP5、LRP6),WNT信號通路調(diào)控器(DVL2、DVL3、GSK-3β, β-CATENIN、APC,AXIN2),WNT信號通路下游轉(zhuǎn)錄因子(TCF4、LEF1)以及WNT信號通路拮抗物(SOSTDC1)的表達(dá)模式,為闡明經(jīng)典WNT信號通路調(diào)控人肺器官發(fā)育機(jī)制奠定基礎(chǔ)。首先Real-time PCR結(jié)果顯示,經(jīng)典WNT信號通路關(guān)鍵基因在人胎肺發(fā)育4個(gè)時(shí)期(7W、12W、17W和21W)均有表達(dá),其中WNT2.D VL2、GSK-3β,β-CATENIN,APC、TCF4及LEF1從7W到12W表達(dá)量逐漸下調(diào),從12W到17W開始升高,但在21W又顯著下調(diào)；LRP5、LRP6、DVL3、AXIN2和SOSTDC1從7W、12W到17W表達(dá)量逐漸升高,但到21W又顯著下調(diào)；WNT7B和FZD7從7W到12W表達(dá)量升高,但從12W到21W又逐漸下降。其次原位雜交結(jié)果顯示,除了經(jīng)典WNT信號通路調(diào)控器DVL3、β-CATENIN, AXIN2及下游轉(zhuǎn)錄因子TCF4在胎肺近端氣管及遠(yuǎn)端肺泡上皮和間充質(zhì)中均有表達(dá)外,其余經(jīng)典WNT信號通路調(diào)控器僅在胎月市氣管分支及遠(yuǎn)端肺泡上皮表達(dá)。此外,我們利用小分子化合物CHIR-99021刺激15周的人胎肺組織以激活經(jīng)典WNT信號通路,Western Blot、Real-time PCR及組織病理檢測證實(shí)經(jīng)典WNT信號通路被激活,并且過度激活的WNT信號致使人胎肺近遠(yuǎn)端肺部上皮細(xì)胞由矮柱狀向高柱狀的早期細(xì)胞狀態(tài)退化。總之,我們的研究表明,經(jīng)典WNT信號通路對人胎肺氣管的分支形態(tài)及上皮細(xì)胞的分化均起著重要的調(diào)控作用,這將為研究經(jīng)典WNT信號通路調(diào)控人胎肺發(fā)育的分子機(jī)制及相關(guān)肺部疾病的診療奠定基礎(chǔ)。
[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.
【學(xué)位授予單位】：福建師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R321

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本文編號：1914515