幽门螺杆菌毒素相关蛋白CagA通过microRNA促进胃上皮细胞间质转化的研究
发布时间：2018-11-06 09:36
【摘要】：实验目的： 探讨幽门螺杆菌(Hp)毒素相关蛋白(CagA)在胃上皮细胞间质转化(Epithelial-mesenchymal transition, EMT)发生发展中的致病机制。 实验方法： 首先建立稳定表达CagA的AGS细胞系,然后分别对CagA转化的胃癌AGS细胞和空载体对照组细胞用哺乳动物miRNA表达谱芯片检测以筛选可能改变的microRNA (miRNA);用荧光虫酶报告基因方法和western blot检测相应miRNA共同作用的重要靶分子；沉默靶分子观察对胃上皮细胞EMT和干细胞比例的影响。 结果： 发现hsa-mir-584和-1290在CagA转化细胞中表达出现了上调,先锋转录因子FoxAl是hsa-mir-584和-1290共同作用的重要靶分子,沉默FoxAl的SW620细胞CD44+CD133+细胞百分比出现明显减少,而CD24+细胞百分比明显增加,P0.05。FoxAl沉默细胞形态由梭型变为小圆型,western blot检测发现E-cadherin表达下降,Vimintin表达增加。 结论： hsa-mir-584和-1290通过下调FoxAl促进了细胞的EMT过程和干扰了细胞的发育和分化,通过miRNA途径是CagA蛋白新的致病机制。
[Abstract]:Objective: to investigate the pathogenetic mechanism of Helicobacter pylori (Hp) toxin associated protein (CagA) in the interstitial transformation of gastric epithelial cells (Epithelial-mesenchymal transition, EMT). Methods: first, AGS cell lines expressing CagA stably were established, then CagA transformed gastric cancer AGS cells and empty vector control cells were detected by mammalian miRNA expression microarray to screen possible microRNA (miRNA); changes. The important target molecules of corresponding miRNA interaction were detected by fluorescent insectase reporter gene method and western blot. The effect of silencing target molecule on EMT and stem cell ratio of gastric epithelial cells was observed. Results: the expression of hsa-mir-584 and-1290 was up-regulated in CagA transformed cells. The pioneer transcription factor FoxAl was an important target molecule of hsa-mir-584 and-1290. The percentage of CD44 CD133 cells in SW620 cells of silencing FoxAl decreased obviously, while the percentage of CD24 cells increased obviously. The morphology of P0.05.FoxAl silencing cells changed from spindle type to small round, western blot. The expression of E-cadherin decreased and Vimintin expression increased. Conclusion: hsa-mir-584 and-1290 promote the EMT process and interfere with the development and differentiation of cells by down-regulating FoxAl. MiRNA pathway is a new pathogenetic mechanism of CagA protein.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R378
本文编号：2313907
[Abstract]:Objective: to investigate the pathogenetic mechanism of Helicobacter pylori (Hp) toxin associated protein (CagA) in the interstitial transformation of gastric epithelial cells (Epithelial-mesenchymal transition, EMT). Methods: first, AGS cell lines expressing CagA stably were established, then CagA transformed gastric cancer AGS cells and empty vector control cells were detected by mammalian miRNA expression microarray to screen possible microRNA (miRNA); changes. The important target molecules of corresponding miRNA interaction were detected by fluorescent insectase reporter gene method and western blot. The effect of silencing target molecule on EMT and stem cell ratio of gastric epithelial cells was observed. Results: the expression of hsa-mir-584 and-1290 was up-regulated in CagA transformed cells. The pioneer transcription factor FoxAl was an important target molecule of hsa-mir-584 and-1290. The percentage of CD44 CD133 cells in SW620 cells of silencing FoxAl decreased obviously, while the percentage of CD24 cells increased obviously. The morphology of P0.05.FoxAl silencing cells changed from spindle type to small round, western blot. The expression of E-cadherin decreased and Vimintin expression increased. Conclusion: hsa-mir-584 and-1290 promote the EMT process and interfere with the development and differentiation of cells by down-regulating FoxAl. MiRNA pathway is a new pathogenetic mechanism of CagA protein.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R378
【参考文献】
相关期刊论文 前1条
1 胡伏莲;;幽门螺杆菌感染与上胃肠道疾病[J];中国医刊;2007年02期
，本文编号：2313907
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