發(fā)布時間：2018-11-06 09:36

【摘要】：實(shí)驗(yàn)?zāi)康模?探討幽門螺桿菌(Hp)毒素相關(guān)蛋白(CagA)在胃上皮細(xì)胞間質(zhì)轉(zhuǎn)化(Epithelial-mesenchymal transition, EMT)發(fā)生發(fā)展中的致病機(jī)制。 實(shí)驗(yàn)方法： 首先建立穩(wěn)定表達(dá)CagA的AGS細(xì)胞系,然后分別對CagA轉(zhuǎn)化的胃癌AGS細(xì)胞和空載體對照組細(xì)胞用哺乳動物miRNA表達(dá)譜芯片檢測以篩選可能改變的microRNA (miRNA);用熒光蟲酶報告基因方法和western blot檢測相應(yīng)miRNA共同作用的重要靶分子；沉默靶分子觀察對胃上皮細(xì)胞EMT和干細(xì)胞比例的影響。 結(jié)果： 發(fā)現(xiàn)hsa-mir-584和-1290在CagA轉(zhuǎn)化細(xì)胞中表達(dá)出現(xiàn)了上調(diào),先鋒轉(zhuǎn)錄因子FoxAl是hsa-mir-584和-1290共同作用的重要靶分子,沉默F(xiàn)oxAl的SW620細(xì)胞CD44+CD133+細(xì)胞百分比出現(xiàn)明顯減少,而CD24+細(xì)胞百分比明顯增加,P0.05。FoxAl沉默細(xì)胞形態(tài)由梭型變?yōu)樾A型,western blot檢測發(fā)現(xiàn)E-cadherin表達(dá)下降,Vimintin表達(dá)增加。 結(jié)論： hsa-mir-584和-1290通過下調(diào)FoxAl促進(jìn)了細(xì)胞的EMT過程和干擾了細(xì)胞的發(fā)育和分化,通過miRNA途徑是CagA蛋白新的致病機(jī)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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R378

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 胡伏蓮;;幽門螺桿菌感染與上胃腸道疾病[J];中國醫(yī)刊;2007年02期

，

本文編號：2313907