肝再生磷酸酶3对胶质瘤细胞增殖和迁移能力的影响

发布时间：2018-04-28 14:42

本文选题：肝再生磷酸酶3 + 脑胶质瘤细胞　；参考：《河南大学》2014年硕士论文

【摘要】：背景：目前研究显示，肝再生磷酸酶3(PRL3)对多种肿瘤细胞的存活、增殖、侵袭、转移具有促进作用，但是具体机制不详，尤其是该分子在脑胶质瘤细胞的增殖和转移中发挥的作用及其机制未见阐明。此文是关于PRL3与脑胶质瘤细胞相关的一些研究。目的：筛选PRL3在细胞中相结合的蛋白，为进一步研究PRL3在信号通路中的作用分子机制提供支持。研究PRL3基因过表达和沉默对脑胶质瘤细胞增殖和迁移能力的影响。方法：pGEX-6P-1-PRL3原核表达载体，诱导GST-PRL3融合蛋白表达并纯化该蛋白，利用pull-down技术筛选小鼠神经细胞中与PRL3相互作用的分子，质谱鉴定与PRL3相互作用蛋白。构建重组pcDNA3.1-PRL3真核表达载体，建立PRL3基因过表达体系，用pcDNA3.1作对照；同时设计并合成siRNA，构建siRNA-pSilencer3.1重组质粒，建立PRL3基因沉默体系，，用pSilencer3.1作对照，四组质粒分别瞬时转染脑胶质瘤U251细胞，经细胞计数、划痕分析等实验观察细胞的增殖和迁移能力。结果：（1）pull-down结果显示，在小鼠脑组织裂解液中有四个未知蛋白条带，质谱鉴定出新的与PRL3相互作用的分子。（2）与对照组相比，PRL3基因过表达的脑胶质瘤U251细胞数量增多，迁移加快；而PRL3基因沉默后脑胶质瘤细胞数量减少，但迁移无明显变化。结论：在小鼠脑组织中筛选出了4种与PRL3相互作用的分子，其中一种经鉴定为Tubulin-beta。PRL3瞬时过表达促进脑胶质瘤细胞增殖和迁移，PRL3沉默抑制脑胶质瘤细胞增殖，但对其迁移没有显著影响。
[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.
【学位授予单位】：河南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41

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