Nanog在胃癌上皮间质转化过程中的作用研究

发布时间：2018-03-27 10:01

本文选题：胃癌　切入点：上皮间质转化　出处：《华北理工大学》2016年硕士论文

【摘要】：目的本文旨在研究多能干性因子Nanog在胃癌上皮间质转化(Epithelialmesenchymal Transition,EMT)过程中的作用及其对胃癌细胞迁移和侵袭能力的影响,并分析Nanog对组蛋白赖氨酸去甲基化酶KDM5B表达的调控,为阐明胃癌发生发展的机制提供基础依据。方法运用免疫组织化学的方法检测胃癌组织、癌旁组织及淋巴结转移组织中Nanog蛋白、上皮标记物E-cadherin和间质标记物N-cadherin的表达情况;免疫细胞化学法检测TGF-β1诱导MGC-803细胞前后Nanog蛋白、E-cadherin和Ncadherin的表达情况,检测Nanog过表达前后和敲低前后E-cadherin、N-cadherin和KDM5B蛋白表达情况;用实时荧光定量PCR法检测Nanog过表达前后和敲低前后Nanog和KDM5B的的m RNA水平;用Western blot法检测Nanog过表达前后和敲低前后Nanog蛋白水平;HE染色法观察Nanog过表达前后和敲低前后MGC-803细胞的形态学变化;划痕实验和Transwell实验检测Nanog过表达前后和敲低前后MGC-803细胞的迁移和侵袭情况。结果1 E-cadherin在癌组织和淋巴结转移组织中的表达水平显著低于癌旁组织(P0.05,P0.05),N-cadherin和Nanog蛋白在癌组织和淋巴结转移组织中的表达水平显著高于癌旁组织(P0.05,P0.05)。2 TGF-β1诱导MGC-803细胞后,E-cadherin表达水平降低(P0.05),N-cadherin和Nanog蛋白的水平增高(P0.05,P0.05)。3 Nanog过表达后的MGC-803细胞排列疏松呈间质样改变;E-cadherin表达水平显著降低(P0.05);N-cadherin和KDM5B蛋白水平均显著增高(P0.05,P0.05);细胞的迁移和侵袭能力均增强(P0.05,P0.05)。4Nanog敲低后的MGC-80细胞排列规则呈上皮样改变;E-cadherin表达水平显著升高(P0.05);N-cadherin和KDM5B蛋白水平均显著降低(P0.05,P0.05);细胞的迁移和侵袭能力均减弱(P0.05,P0.05)。结论1在发生了EMT的胃腺癌组织和细胞中多能干性因子Nanog呈高表达。2 Nanog可正向调控胃腺癌细胞EMT、迁移和侵袭能力。3胃腺癌细胞中,Nanog正向调控组蛋白赖氨酸去甲基化酶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.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.2

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