本文選題：肝再生磷酸酶3 + 腦膠質(zhì)瘤細(xì)胞��； 參考：《河南大學(xué)》2014年碩士論文

【摘要】：背景：目前研究顯示，肝再生磷酸酶3(PRL3)對(duì)多種腫瘤細(xì)胞的存活、增殖、侵襲、轉(zhuǎn)移具有促進(jìn)作用，但是具體機(jī)制不詳，尤其是該分子在腦膠質(zhì)瘤細(xì)胞的增殖和轉(zhuǎn)移中發(fā)揮的作用及其機(jī)制未見闡明。此文是關(guān)于PRL3與腦膠質(zhì)瘤細(xì)胞相關(guān)的一些研究。 目的：篩選PRL3在細(xì)胞中相結(jié)合的蛋白，為進(jìn)一步研究PRL3在信號(hào)通路中的作用分子機(jī)制提供支持。研究PRL3基因過表達(dá)和沉默對(duì)腦膠質(zhì)瘤細(xì)胞增殖和遷移能力的影響。 方法：pGEX-6P-1-PRL3原核表達(dá)載體，誘導(dǎo)GST-PRL3融合蛋白表達(dá)并純化該蛋白，利用pull-down技術(shù)篩選小鼠神經(jīng)細(xì)胞中與PRL3相互作用的分子，質(zhì)譜鑒定與PRL3相互作用蛋白。構(gòu)建重組pcDNA3.1-PRL3真核表達(dá)載體，建立PRL3基因過表達(dá)體系，用pcDNA3.1作對(duì)照；同時(shí)設(shè)計(jì)并合成siRNA，構(gòu)建siRNA-pSilencer3.1重組質(zhì)粒，建立PRL3基因沉默體系，，用pSilencer3.1作對(duì)照，四組質(zhì)粒分別瞬時(shí)轉(zhuǎn)染腦膠質(zhì)瘤U251細(xì)胞，經(jīng)細(xì)胞計(jì)數(shù)、劃痕分析等實(shí)驗(yàn)觀察細(xì)胞的增殖和遷移能力。 結(jié)果：（1）pull-down結(jié)果顯示，在小鼠腦組織裂解液中有四個(gè)未知蛋白條帶，質(zhì)譜鑒定出新的與PRL3相互作用的分子。（2）與對(duì)照組相比，PRL3基因過表達(dá)的腦膠質(zhì)瘤U251細(xì)胞數(shù)量增多，遷移加快；而PRL3基因沉默后腦膠質(zhì)瘤細(xì)胞數(shù)量減少，但遷移無(wú)明顯變化。 結(jié)論：在小鼠腦組織中篩選出了4種與PRL3相互作用的分子，其中一種經(jīng)鑒定為Tubulin-beta。PRL3瞬時(shí)過表達(dá)促進(jìn)腦膠質(zhì)瘤細(xì)胞增殖和遷移，PRL3沉默抑制腦膠質(zhì)瘤細(xì)胞增殖，但對(duì)其遷移沒有顯著影響。
[Abstract]:Background: current studies have shown that liver regeneration phosphatase 3 (PRL3) can promote the survival, proliferation, invasion and metastasis of various tumor cells, but the specific mechanism is unknown. In particular, the role of this molecule in the proliferation and metastasis of glioma cells and its mechanism have not been elucidated. This article is about PRL3 and glioma cells related to some studies. Aim: to screen PRL3 binding proteins in order to support the further study of the molecular mechanism of PRL3 in signal pathway. To study the effect of overexpression and silencing of PRL3 gene on the proliferation and migration of glioma cells. Methods the prokaryotic expression vector of pGEX-6P-1-PRL3 was used to induce the expression and purification of GST-PRL3 fusion protein. The molecules interacting with PRL3 in mouse nerve cells were screened by pull-down technique. The interaction protein with PRL3 was identified by mass spectrometry. The eukaryotic expression vector of recombinant pcDNA3.1-PRL3 was constructed, the overexpression system of PRL3 gene was established, and pcDNA3.1 was used as control. At the same time, siRNA-pSilencer3.1 recombinant plasmid was constructed, PRL3 gene silencing system was established and pSilencer3.1 was used as control. Four groups of plasmids were transiently transfected into glioma U251 cells. The proliferation and migration of U251 cells were observed by cell count and scratch analysis. Results the results showed that there were four unknown protein bands in the cleavage fluid of mouse brain tissue, and the new molecule of interaction with PRL3 was identified by mass spectrometry. Compared with the control group, the number of U251 cells with overexpression of PRL3 gene was increased and the migration of U251 cells was accelerated. However, the number of glioma cells decreased after PRL3 gene silencing, but there was no significant change in migration. Conclusion: four molecules interacting with PRL3 were screened in mouse brain tissue, one of which was identified as transient overexpression of Tubulin-beta.PRL3 to promote the proliferation and migration of glioma cells to inhibit the proliferation of glioma cells. However, there was no significant effect on its migration.
【學(xué)位授予單位】：河南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.41

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