發(fā)布時間：2018-07-18 20:32

【摘要】：催化N-聯(lián)接聚糖形成的糖基轉(zhuǎn)移酶表達(dá)量的變化與多種細(xì)胞生物學(xué)行為有關(guān)，如：細(xì)胞粘附，細(xì)胞遷移，細(xì)胞增值及腫瘤細(xì)胞轉(zhuǎn)移等。本研究中，我們觀察到了N-乙酰氨基葡萄糖轉(zhuǎn)移酶V(GnT-V)及其催化產(chǎn)物（β1，6-GlcNAc）表達(dá)量的變化與炎癥反應(yīng)有關(guān)：GnT-V參與調(diào)控整合素α5β1介導(dǎo)的單核細(xì)胞-內(nèi)皮細(xì)胞的粘附和遷移。通過細(xì)胞粘附和遷移實(shí)驗(yàn)，我們觀察到IFN-γ所處理的單核細(xì)胞THP-1對血管內(nèi)皮細(xì)胞EA.hy926的粘附和侵襲能力明顯增強(qiáng)；同時，QRT-PCR及westernblot方法檢測到此類單核細(xì)胞內(nèi)的GnT-V及其催化產(chǎn)物（β1,6-GlcNAc）表達(dá)量明顯下降。采用GnT-V干擾質(zhì)粒轉(zhuǎn)染單核細(xì)胞THP-1后，其對血管內(nèi)皮的粘附和遷移能力亦明顯增強(qiáng)，與IFN-γ處理具有相同生物學(xué)效應(yīng)。我們還發(fā)現(xiàn)，通過封閉整合素α5或β1亞基可以逆轉(zhuǎn)GnT-V干擾及IFN-γ處理所增強(qiáng)的單核細(xì)胞粘附和侵襲能力；westernblot方法驗(yàn)證證明上述處理減少整合素α5或β1亞基表面β1,6-GlcNAc的糖鏈修飾，而并不影響單核細(xì)胞內(nèi)整合素α5或β1亞基表達(dá)。在相關(guān)信號通路的研究中發(fā)現(xiàn)，GnT-V表達(dá)量的下降明顯增強(qiáng)整合素介導(dǎo)的FAK磷酸化。同時，我們觀察到FAK磷酸化增強(qiáng)可激活其下游信號通路-ERK，而ERK磷酸化抑制劑預(yù)處理IFN-γ刺激的單核細(xì)胞后其對內(nèi)皮的黏附和侵襲能力無明顯增強(qiáng)。綜上，我們的研究第一次發(fā)現(xiàn)炎癥因子處理單核細(xì)胞所誘導(dǎo)的炎癥反應(yīng)中，GnT-V活性明顯下降，從而增強(qiáng)整合素α5β1介導(dǎo)的單核細(xì)胞-血管內(nèi)皮黏附和遷移；同時推測整合素-FAK信號通路及FAK信號通路下游-ERK信號通路可能參與上述生物學(xué)效應(yīng)的調(diào)控。GnT-V可能成為心腦血管炎癥疾病研究及治療的新靶點(diǎn)。
[Abstract]:The changes in the expression of glycosyltransferases catalyzing the formation of N-ligand are related to a variety of cell biological behaviors, such as cell adhesion, cell migration, cell proliferation and tumor cell metastasis. In this study, we observed that the expression of N-acetylglucosaminotransferase V (GnT-V) and its catalytic product (尾 _ 1N _ (6-GlcNAc) were related to the inflammatory response, and that: GnT-V was involved in regulating the adhesion and migration of integrin 偽 _ 5 尾 _ 1-mediated monocyte to endothelial cells. Through cell adhesion and migration experiments, we observed that the adhesion and invasion ability of THP-1 treated with IFN- 緯 on vascular endothelial cell EA.hy926 was significantly enhanced. At the same time, the expression of GnT-V and its catalytic product (尾 1N 6-GlcNAc) in these monocytes were detected by QRT-PCR and westernblot. After transfection with GnT-V interference plasmid, the adhesion and migration of THP-1 to vascular endothelium were significantly enhanced, which had the same biological effect as IFN- 緯 treatment. We also found that blocking integrin 偽 5 or 尾 1 subunit could reverse GnT-V interference and the enhanced adhesion and invasion ability of monocytes treated with IFN- 緯. Western blot showed that these treatments reduced the sugar chain modification of integrin 偽 5 or 尾 1 subunit surface 尾 1N 6-GlcNAc. The expression of integrin 偽 5 or 尾 1 subunit in monocytes was not affected. It was found that the decrease of GnT-V expression significantly enhanced integrin-mediated FAK phosphorylation. At the same time, we observed that enhanced phosphorylation of FAK activated its downstream signaling pathway -ERK, but the adhesion and invasion of IFN- 緯 stimulated monocytes were not significantly enhanced after pretreatment with ERK phosphorylation inhibitor. In conclusion, we found for the first time that the activity of GnT-V decreased significantly in the inflammatory response induced by inflammatory cytokines, which enhanced the adhesion and migration of integrin 偽 5 尾 1-mediated monocytes to vascular endothelium. It is also speculated that integrin-FAK signaling pathway and its downstream -ERK signaling pathway may be involved in the regulation of these biological effects. GnT-V may be a new target for the study and treatment of cardiovascular and cerebrovascular inflammatory diseases.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

【共引文獻(xiàn)】

相關(guān)期刊論文 前3條

1 張婷;蔣春雷;;腫瘤條件培養(yǎng)基對人臍靜脈內(nèi)皮細(xì)胞增殖、黏附、遷移能力的調(diào)節(jié)[J];生理學(xué)報(bào);2011年03期

2 徐統(tǒng)震;孫雪飛;任冬梅;楊國濤;;木犀草素抑制肺癌細(xì)胞A549的增殖及其聯(lián)合化療作用[J];山東大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2012年07期

3 楊火梅;于超;楊竹;;功能蛋白的O-糖基化和p38磷酸化共同參與調(diào)控單核細(xì)胞對血管內(nèi)皮的粘附和侵襲[J];中國細(xì)胞生物學(xué)學(xué)報(bào);2012年03期

相關(guān)博士學(xué)位論文 前2條

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2 周峰;E-鈣粘蛋白Asn-633位N-糖鏈對其表達(dá)、轉(zhuǎn)運(yùn)和折疊的影響及分子機(jī)制的研究[D];復(fù)旦大學(xué);2008年

相關(guān)碩士學(xué)位論文 前2條

1 徐良;唾液酸轉(zhuǎn)移酶ST8SiaⅥ在乳腺癌細(xì)胞轉(zhuǎn)移過程中的作用及其分子機(jī)制的初步研究[D];中國海洋大學(xué);2010年

2 李亞輝;沉默N-乙酰糖基轉(zhuǎn)移酶V抑制CNE-2細(xì)胞的增殖和轉(zhuǎn)移能力[D];南方醫(yī)科大學(xué);2011年

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本文編號：2132688