炎症因子在慢性砷诱导永生化人肺支气管上皮细胞转化的作用

发布时间：2018-06-12 11:45

本文选题：肺支气管上皮BEAS-2B细胞 + 亚砷酸钠　；参考：《福建医科大学》2014年硕士论文

【摘要】：目的：砷污染已经成为全世界范围内受到广泛关注的环境问题之一，慢性砷中毒对人体多器官系统功能均可造成危害，并引起肝癌、肾癌、肺癌、膀胱癌和皮肤癌。皮间质转化（Epithelial-mesenchymal Transition）与肿瘤细胞的生长、侵袭、迁移有重要关联。炎症在肿瘤微环境中有促肿瘤的作用，促进恶性细胞的生存和增殖。本文旨在探讨炎症因子在慢性砷诱导永生化人肺支气管上皮细胞转化的作用。方法： 1.永生化人肺支气管上皮细胞BEAS-2B经0.25uM亚砷酸钠暴露培养66天，并进行软琼脂集落形成实验，挑取细胞克隆消化培养为转化细胞；对照组BEAS-2B相同条件无亚砷酸钠培养66天。 2. Westernblot检测上皮细胞标志蛋白E-Cadherin、间质细胞标志蛋白Vimentin表达。 3. Westernblot检测炎性体蛋白AIM2、ASC、炎性体通路下游炎症细胞因子IL-1β 4. Westernblot检测炎症因子HMGB1的表达。结果： 1. BEAS-2B细胞经慢性亚砷酸钠暴露66天，克隆形成率为22.77%，，较对照组（3.06%）具有显著差异（p0.01）。 2.慢性砷诱导转化的BEAS-2B细胞，形态由上皮细胞的不规则多边形变为梭形。 3.转化的BEAS-2B细胞较对照组上皮细胞标志蛋白E-Cadherin表达减少、间质细胞标志蛋白Vimentin表达增加。 4.转化的BEAS-2B细胞较对照组炎性体蛋白AIM2、ASC表达减少，炎性体通路下游炎症细胞因子IL-1β表达减少。 5.转化的BEAS-2B细胞HMGB1表达增加。结论： 1.慢性砷诱导的BEAS-2B细胞形态发生改变，克隆形成能力增强，细胞发生转化。 2.炎性体蛋白表达减少在BEAS-2B转化中起一定的促进作用。 3.转化的BEAS-2B细胞HMGB1表达增加，提示细胞转化与HMGB1表达增强有关。
[Abstract]:Objective: arsenic pollution has become one of the most important environmental problems all over the world. Chronic arsenic poisoning is harmful to the function of human multi-organ system, and can cause liver cancer, kidney cancer, lung cancer, bladder cancer and skin cancer. Epithelial-mesenchymal Transitionis associated with the growth, invasion and migration of tumor cells. Inflammation promotes the survival and proliferation of malignant cells in tumor microenvironment. The aim of this study was to investigate the role of inflammatory factors in chronic arsenic induced transformation of immortalized human lung bronchial epithelial cells. The immortalized human lung bronchial epithelial cells BEAS-2B were exposed to 0.25 UM sodium arsenite for 66 days, and the soft Agar colony formation test was carried out to select the cells for clone digestion and culture into transformed cells, while the control group BEAS-2B was cultured without sodium arsenite for 66 days under the same conditions. 2. The expression of E-Cadherin in epithelial cells and Vimentin in interstitial cells was detected by Western blot. Western blot was used to detect the inflammatory protein AIM2 and the inflammatory cytokine IL-1 尾 4 downstream of the inflammatory pathway. The expression of HMGB1 was detected by Western blot. Results: 1. When BEAS-2B cells were exposed to chronic sodium arsenite for 66 days, the clone formation rate was 22.77, which was significantly different from that of the control group (3.06). The morphology of BEAS-2B cells induced by chronic arsenic was changed from irregular polygon of epithelial cells to fusiform. The expression of E-Cadherin in the transformed BEAS-2B cells was lower than that in the control group, and the expression of Vimentin was increased in the stromal cells. Compared with the control group, the expression of AIM2 + ASC in BEAS-2B cells was lower than that in the control group, and the expression of IL-1 尾 was decreased in the downstream of inflammatory pathway. The expression of HMGB1 in transformed BEAS-2B cells was increased. Conclusion: 1. BEAS-2B cells induced by chronic arsenic had morphologic changes, enhanced clone formation and cell transformation. 2. 2. The decrease of inflammatory body protein expression in BEAS-2B transformation plays a certain role in promoting. 3. The expression of HMGB1 in transformed BEAS-2B cells was increased, which suggested that the expression of HMGB1 was related to the expression of HMGB1 in transformed BEAS-2B cells.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R114

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