本文選題：非小細(xì)胞肺癌 + 肺腺癌��； 參考：《吉林大學(xué)》2017年博士論文

【摘要】：肺癌是一種致死性、惡性疾病,因其發(fā)病率逐年增加、預(yù)后差,是嚴(yán)重威脅人類的公共健康問題。雖然有大量針對(duì)肺癌的分子生物學(xué)實(shí)驗(yàn),但我們對(duì)肺癌的發(fā)生、發(fā)展中的分子生物學(xué)機(jī)制仍不十分清楚。sprouty1是已知的RTK信號(hào)轉(zhuǎn)導(dǎo)通路的抑制劑,可以通過調(diào)節(jié)EGF、FGF、PDGF等多種途徑介導(dǎo)的信號(hào)轉(zhuǎn)導(dǎo)通路影響細(xì)胞的增殖、分化、轉(zhuǎn)移。鑒于國(guó)內(nèi)外尚沒有sprouty1在肺腺癌中的表達(dá)及其作用機(jī)制的相關(guān)報(bào)道,我們首先構(gòu)建sprouty1基因敲除小鼠模型,利用免疫組織化學(xué)染色和免疫熒光染色的實(shí)驗(yàn)方法,驗(yàn)證sprouty1在肺組織中的表達(dá)情況,觀察sprouty1基因敲除后對(duì)小鼠肺組織表型的影響、以及對(duì)Ⅱ型肺泡上皮細(xì)胞功能的影響。其次,通過HE染色、免疫組化染色的方法比較sprouty1基因敲除小鼠與野生型小鼠的肺癌發(fā)病率的差異,以及兩種基因型小鼠肺組織中TTF1的表達(dá)情況。而且將在體外利用細(xì)胞轉(zhuǎn)染技術(shù)構(gòu)建sprouty1過表達(dá)的A549細(xì)胞系,驗(yàn)證sprouty1對(duì)肺癌細(xì)胞增殖、分化的影響。最后,利用免疫熒光染色、western-blot等方法進(jìn)一步探討sprouty1在肺腺癌發(fā)生、發(fā)展中的生物學(xué)功能,以及可能的作用機(jī)制,最終揭示sprouty1的抑癌作用本質(zhì)。通過上述實(shí)驗(yàn),我們得到以下結(jié)果:1、sprouty1在肺組織中高度表達(dá),主要是由Ⅱ型肺泡上皮細(xì)胞(alveolar epithelial cell typeⅡ,AECⅡ)表達(dá),sprouty1基因敲除后導(dǎo)致AECⅡ表達(dá)PCNA和Ki-67增加,說明sprouty1基因敲除后促進(jìn)Ⅱ型肺泡上皮細(xì)胞的增殖。2、sprouty1基因敲除小鼠較野生型小鼠的肺癌發(fā)生率明顯增加(P0.01),且腫瘤的病理學(xué)分型符合肺腺癌。3、體外過表達(dá)sprouty1是A549細(xì)胞的細(xì)胞周期相關(guān)蛋白表達(dá)減少,說明sprouty1可抑制肺腺癌細(xì)胞的增殖。4、sprouty1基因敲除后導(dǎo)致小鼠肺組織中EGFR、FGFR2和p ERK表達(dá)增加,且p ERK與PCNA存在共染色,說明sprouty是通過EGF、FGF介導(dǎo)的ERK信號(hào)轉(zhuǎn)導(dǎo)通路調(diào)控細(xì)胞的增殖。5、sprouty1基因敲除后小鼠肺組織中vimentin、β1-integrin、SMA、PDGFRβ和fibronectin的蛋白表達(dá)增加,說明sprouty1可通過調(diào)控細(xì)胞的EMT抑制肺腺癌的侵襲。通過以上實(shí)驗(yàn)結(jié)果,我們認(rèn)為sprouty1可被認(rèn)為是一種肺腺癌的抑癌基因,主要通過抑制EGF、FGF介導(dǎo)的Ras/ERK信號(hào)轉(zhuǎn)導(dǎo)通路和抑制腺癌細(xì)胞的EMT這兩大機(jī)制,從而抑制肺腺癌細(xì)胞的增殖和侵襲。未來,sprouty1也許可以成為肺癌治療的新靶點(diǎn)。
[Abstract]:Lung cancer is a fatal and malignant disease. Because of its increasing incidence and poor prognosis, lung cancer is a serious public health problem. Although there are a lot of molecular biological experiments aimed at lung cancer, we do not know that the molecular biological mechanism of lung cancer is very clear that .sprouty1 is an inhibitor of known RTK signal transduction pathway. It can affect the proliferation, differentiation and metastasis of cells by regulating the signal transduction pathway mediated by EGFG FGF- PDGF and other pathways. Since there are no reports on the expression and mechanism of sprouty1 in lung adenocarcinoma at home and abroad, we first constructed the sprouty1 gene knockout mouse model and used immunohistochemical staining and immunofluorescence staining methods. To verify the expression of sprouty1 in lung tissue and observe the effect of sprouty1 gene knockout on the phenotype of lung tissue and the function of type 鈪，

本文編號(hào)：1861588