本文選題：膠質(zhì)瘤 + 星形膠質(zhì)細(xì)胞　； 參考：《福建醫(yī)科大學(xué)》2014年博士論文

【摘要】：目的研究顯示膠質(zhì)瘤細(xì)胞可通過釋放大量的谷氨酸來促進(jìn)自身生長、殺傷神經(jīng)元，而星形膠質(zhì)細(xì)胞是處理谷氨酸的主要載體，通過研究星形膠質(zhì)細(xì)胞與膠質(zhì)瘤細(xì)胞之間的相互作用，探討星形膠質(zhì)細(xì)胞在限制膠質(zhì)瘤細(xì)胞所釋放的胞外谷氨酸水平的作用，揭示內(nèi)源性抑制膠質(zhì)瘤生長及保護(hù)神經(jīng)元的機(jī)制。為了盡量避免因現(xiàn)有的細(xì)胞株傳代久遠(yuǎn)因素引起的實(shí)驗(yàn)結(jié)果的局限性，，本研究將建立膠質(zhì)瘤細(xì)胞株，并利用傳代10代以內(nèi)的膠質(zhì)瘤細(xì)胞進(jìn)行實(shí)驗(yàn)。 方法利用細(xì)胞培養(yǎng)技術(shù)、細(xì)胞示蹤及凋亡檢測、免疫細(xì)胞染色技術(shù)、westernblot技術(shù)、藥物干預(yù)、高效液相色譜檢測谷氨酸和谷氨酰胺、離子成像等技術(shù)，模擬體內(nèi)膠質(zhì)瘤生長過程，驗(yàn)證如下假說：1、在膠質(zhì)瘤生長的早期，其周圍包繞大量正常的星形膠質(zhì)細(xì)胞，此時(shí)（1）膠質(zhì)瘤細(xì)胞是否高表達(dá)谷氨酰胺酶，而不表達(dá)或錯(cuò)位表達(dá)與谷氨酰胺酶作用相反的谷氨酰胺合成酶；（2）膠質(zhì)瘤細(xì)胞能否大量攝取谷氨酰胺進(jìn)入胞內(nèi)并轉(zhuǎn)化為谷氨酸分泌到細(xì)胞外；（3）星形膠質(zhì)細(xì)胞能否有效地?cái)z取膠質(zhì)瘤細(xì)胞所分泌的谷氨酸；2、在膠質(zhì)瘤生長的后期，其數(shù)量上超過周圍包繞的星形膠質(zhì)細(xì)胞，此狀態(tài)下谷氨酸-谷氨酰胺代謝耦合是否被打破：（1）膠質(zhì)瘤細(xì)胞所釋放的氨及氧化應(yīng)激能否在一定程度上抑制星形膠質(zhì)細(xì)胞的谷氨酸攝取功能，而使共培養(yǎng)中胞外谷氨酸水平明顯升高，引起神經(jīng)元的損傷；（2）大量的膠質(zhì)瘤細(xì)胞能否通過氧化應(yīng)激等造成星形膠質(zhì)細(xì)胞的損傷甚至死亡。 結(jié)果根據(jù)以下特征（1）膠質(zhì)瘤細(xì)胞高表達(dá)谷氨酰胺酶，但谷氨酰胺合成酶錯(cuò)位表達(dá)。而星形膠質(zhì)細(xì)胞高表達(dá)谷氨酰胺合成酶；（2）膠質(zhì)瘤細(xì)胞能夠大量攝取谷氨酰胺進(jìn)入細(xì)胞內(nèi)并轉(zhuǎn)化成為谷氨酸，再主要通過胱氨酸-谷氨酸交換蛋白分泌到細(xì)胞外；（3）在星形膠質(zhì)細(xì)胞與膠質(zhì)瘤細(xì)胞共培養(yǎng)過程中，星形膠質(zhì)細(xì)胞能夠有效攝取膠質(zhì)瘤細(xì)胞所分泌的谷氨酸，證明星形膠質(zhì)細(xì)胞與膠質(zhì)瘤細(xì)胞之間存在谷氨酸-谷氨酰胺代謝耦合。而在膠質(zhì)瘤生長的后期，膠質(zhì)瘤體積增大，數(shù)量增多，此狀態(tài)下谷氨酸-谷氨酰胺代謝耦合被打破：（1）膠質(zhì)瘤細(xì)胞所釋放的氨及氧化應(yīng)激顯著抑制星形膠質(zhì)細(xì)胞的谷氨酸攝取功能，而使共培養(yǎng)中的胞外谷氨酸水平明顯升高；（2）胞外谷氨酸明顯升高到能夠誘導(dǎo)神經(jīng)元的損傷及死亡；（3）膠質(zhì)瘤細(xì)胞所產(chǎn)生的大量氨和氧化應(yīng)激還能明顯誘導(dǎo)星形膠質(zhì)細(xì)胞凋亡。 結(jié)論在膠質(zhì)瘤生長的早期，膠質(zhì)瘤細(xì)胞與星形膠質(zhì)細(xì)胞存在谷氨酸-谷氨酰胺代謝耦合，而隨著膠質(zhì)瘤的生長，谷氨酸-谷氨酰胺代謝耦合逐漸被膠質(zhì)瘤的谷氨酸釋放所主導(dǎo)，造成神經(jīng)元和星形膠質(zhì)細(xì)胞的損傷和死亡，進(jìn)一步為膠質(zhì)瘤的生長和侵襲提供空間，同時(shí)胞外升高的谷氨酸可能進(jìn)一步促進(jìn)膠質(zhì)瘤細(xì)胞的生長，使膠質(zhì)瘤進(jìn)入快速生長階段。因此，保持星形膠質(zhì)細(xì)胞與膠質(zhì)瘤細(xì)胞之間谷氨酸-谷氨酰胺代謝耦合的平衡以及維持星形膠質(zhì)細(xì)胞的主導(dǎo)作用，可能可以作為內(nèi)源性抗腫瘤生長和保護(hù)神經(jīng)元的機(jī)制。
[Abstract]:Objective to show that glioma cells can promote their growth and kill neurons by releasing a large amount of glutamic acid, and astrocytes are the main carrier of glutamate treatment. By studying the interaction between astrocytes and glioma cells, astrocytes are used to explore the extracellular valley of astrocytes in the restriction of glioma cells. In order to avoid the limitations of the experimental results caused by the long distant factors of the existing cell lines, this study will establish glioma cell lines and make use of glioma cells within 10 generations to carry out experiments.
Methods using cell culture technique, cell tracer and apoptosis detection, immuno cell staining technique, Westernblot technology, drug intervention, high performance liquid chromatography to detect glutamic acid and glutamine, ion imaging techniques to simulate the growth process of glioma in vivo, verify the following hypothesis: 1, around the early growth of glioma, the surrounding package around a large number of normal. Astrocytes, at this time (1) whether glioma cells are highly expressed as glutaminase, and do not express or misrepresent glutamine synthetase opposite to glutaminase; (2) whether glioma cells can absorb glutamine into the cell and convert into glutamic acid out of the cell; (3) whether astrocytes are effective or not. Uptake of glutamate secreted by glioma cells; 2, in the late stage of the growth of glioma, the number exceeds the surrounding astrocytes. In this state, whether the glutamic glutamine metabolic coupling is broken: (1) whether the ammonia and oxidative stress released by glioma cells can inhibit glutamic acid to a certain extent The level of extracellular glutamic acid increased significantly in co culture, causing neuronal damage, and (2) whether a large number of glioma cells could cause astrocyte damage or even death through oxidative stress.
Results according to the following characteristics (1) glioma cells expressed glutaminase high, but glutamine synthetase was misplaced, and astrocytes highly expressed glutamine synthetase; (2) glioma cells were able to absorb glutamine into cells and convert into glutamic acid, and then secreted mainly through cystine glutamic acid exchange protein. (3) in the process of co culture of astrocytes and glioma cells, astrocytes can effectively absorb glutamic acid secreted by glioma cells and prove that there is a metabolic coupling between glutamate and glutamine in astrocytes and glioma cells. In the later period of the growth of gelatoma, the volume of glioma increases and the number of glioma is increased. In addition, the metabolic coupling of glutamic glutamine was broken in this state: (1) the ammonia and oxidative stress released by glioma cells significantly inhibited the glutamate uptake of astrocytes and increased the level of extracellular glutamic acid in co culture; (2) extracellular glutamic acid significantly increased to the ability to induce neuronal damage and death. (3) a large amount of ammonia and oxidative stress produced by glioma cells can also significantly induce astrocyte apoptosis.
Conclusion in the early stage of glioma growth, glioma cells and astrocytes are coupled with glutamic glutamine metabolism, and with the growth of glioma, the metabolic coupling of glutamic acid is gradually dominated by the release of glutamic acid in glioma, causing damage and death of neurons and astrocytes, and further to glioma. Growth and invasion provide space, while extracellular elevated glutamic acid may further promote the growth of glioma cells and make glioma into the rapid growth stage. Therefore, it is possible to maintain the balance of glutamate metabolic coupling between astrocytes and glioma cells and the dominant role of maintaining astrocytes in astrocytes. As an endogenous mechanism of anti-tumor growth and protection of neurons.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R739.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 孫洪贊;范國光;王桂玲;郭啟勇;;腦膠質(zhì)瘤致癲沲的化學(xué)突觸機(jī)制研究進(jìn)展[J];北京大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2008年04期

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