本文選題：胃癌　切入點(diǎn)：上皮間質(zhì)轉(zhuǎn)化　出處：《華北理工大學(xué)》2016年碩士論文

【摘要】：目的本文旨在研究多能干性因子Nanog在胃癌上皮間質(zhì)轉(zhuǎn)化(Epithelialmesenchymal Transition,EMT)過程中的作用及其對胃癌細(xì)胞遷移和侵襲能力的影響,并分析Nanog對組蛋白賴氨酸去甲基化酶KDM5B表達(dá)的調(diào)控,為闡明胃癌發(fā)生發(fā)展的機(jī)制提供基礎(chǔ)依據(jù)。方法運(yùn)用免疫組織化學(xué)的方法檢測胃癌組織、癌旁組織及淋巴結(jié)轉(zhuǎn)移組織中Nanog蛋白、上皮標(biāo)記物E-cadherin和間質(zhì)標(biāo)記物N-cadherin的表達(dá)情況;免疫細(xì)胞化學(xué)法檢測TGF-β1誘導(dǎo)MGC-803細(xì)胞前后Nanog蛋白、E-cadherin和Ncadherin的表達(dá)情況,檢測Nanog過表達(dá)前后和敲低前后E-cadherin、N-cadherin和KDM5B蛋白表達(dá)情況;用實(shí)時(shí)熒光定量PCR法檢測Nanog過表達(dá)前后和敲低前后Nanog和KDM5B的的m RNA水平;用Western blot法檢測Nanog過表達(dá)前后和敲低前后Nanog蛋白水平;HE染色法觀察Nanog過表達(dá)前后和敲低前后MGC-803細(xì)胞的形態(tài)學(xué)變化;劃痕實(shí)驗(yàn)和Transwell實(shí)驗(yàn)檢測Nanog過表達(dá)前后和敲低前后MGC-803細(xì)胞的遷移和侵襲情況。結(jié)果1 E-cadherin在癌組織和淋巴結(jié)轉(zhuǎn)移組織中的表達(dá)水平顯著低于癌旁組織(P0.05,P0.05),N-cadherin和Nanog蛋白在癌組織和淋巴結(jié)轉(zhuǎn)移組織中的表達(dá)水平顯著高于癌旁組織(P0.05,P0.05)。2 TGF-β1誘導(dǎo)MGC-803細(xì)胞后,E-cadherin表達(dá)水平降低(P0.05),N-cadherin和Nanog蛋白的水平增高(P0.05,P0.05)。3 Nanog過表達(dá)后的MGC-803細(xì)胞排列疏松呈間質(zhì)樣改變;E-cadherin表達(dá)水平顯著降低(P0.05);N-cadherin和KDM5B蛋白水平均顯著增高(P0.05,P0.05);細(xì)胞的遷移和侵襲能力均增強(qiáng)(P0.05,P0.05)。4Nanog敲低后的MGC-80細(xì)胞排列規(guī)則呈上皮樣改變;E-cadherin表達(dá)水平顯著升高(P0.05);N-cadherin和KDM5B蛋白水平均顯著降低(P0.05,P0.05);細(xì)胞的遷移和侵襲能力均減弱(P0.05,P0.05)。結(jié)論1在發(fā)生了EMT的胃腺癌組織和細(xì)胞中多能干性因子Nanog呈高表達(dá)。2 Nanog可正向調(diào)控胃腺癌細(xì)胞EMT、遷移和侵襲能力。3胃腺癌細(xì)胞中,Nanog正向調(diào)控組蛋白賴氨酸去甲基化酶KDM5B。
[Abstract]:Objective to investigate the role of Nanog in the process of epithelial mesenchymal transition (EMTT) and its effect on the migration and invasion of gastric cancer cells, and to analyze the regulation of Nanog on the expression of histone lysine demethylase (KDM5B). Methods the expression of Nanog protein, epithelial marker E-cadherin and interstitial marker N-cadherin in gastric cancer tissues, paracancerous tissues and lymph node metastasis tissues were detected by immunohistochemical method. The expression of E-cadherin and Ncadherin in MGC-803 cells induced by TGF- 尾 1 was detected by immunocytochemistry, and the expression of E-cadherin and KDM5B protein was detected before and after Nanog overexpression and before and after knock down. The m RNA levels of Nanog and KDM5B before and after Nanog overexpression and before and after knockout were detected by real-time fluorescence quantitative PCR. The levels of Nanog protein before and after Nanog overexpression and before and after knock down were detected by Western blot method. The morphological changes of MGC-803 cells before and after Nanog overexpression were observed by HE staining. The migration and invasion of MGC-803 cells before and after Nanog overexpression and before and after knockout were detected by scratch test and Transwell assay. Results 1 the expression of E-cadherin in cancer tissues and lymph node metastasis tissues was significantly lower than that in adjacent tissues (P0.05, P0.05, N-cadherin and Nanog protein). The expression level of E-cadherin in MGC-803 cells induced by P0.05TGF- 尾 1 was significantly higher than that in adjacent tissues with lymph node metastasis. The levels of E-cadherin and E-cadherin protein in MGC-803 cells induced by P0.05TGF- 尾 1 decreased. The levels of N-cadherin and Nanog proteins increased. The MGC-803 cells arranged loosely and showed interstitial changes after overexpression of P0.05 Nanog. The expression of P0.05 N-cadherin and KDM5B protein were significantly increased, and the migration and invasion ability of P0.05 and P0.05 P0.05G knockout were enhanced, and the MGC-80 cells arranged in the same way as epithelium. The expression level of E-cadherin increased significantly. The expression of P0.05N-cadherin and KDM5B protein increased significantly. The migration and invasion ability of the cells were all decreased. Conclusion 1 the expression of Nanog in gastric adenocarcinoma tissues and cells with EMT is high. 2. 2 Nanog can positively regulate the gastric adenocarcinoma cell line EMTT, and the ability of migration and invasion. 3. 3 gastric gland carcinoma cells. Nanog positively regulated histone lysine demethylase KDM5B in cancer cells.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R735.2

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本文編號：1671070