HAUSP表达水平对胶质瘤细胞增殖和凋亡的影响

发布时间：2018-04-09 13:20

  本文选题：神经胶质瘤　切入点：HAUSP　出处：《华中科技大学学报(医学版)》2017年06期

【摘要】：目的探讨调控疱疹病毒相关泛素特异性蛋白酶(HAUSP)对胶质瘤细胞增殖和凋亡的影响。方法构建HAUSP过表达及低表达质粒,转染胶质瘤U87、U251细胞,上调或下调胶质瘤细胞中HAUSP表达水平。荧光定量PCR、Western blot检测胶质瘤细胞中HAUSP mRNA和蛋白表达水平;利用四甲基偶氮唑盐比色法(MTT)检测细胞增殖水平;流式细胞术检测细胞凋亡变化。同时检测各组细胞NANOG mRNA和蛋白表达情况。结果成功构建稳定HAUSP过表达和低表达U87、U251细胞株;与空白对照组相比,HAUSP过表达组细胞中,HAUSP的mRNA、蛋白水平均明显增加,NANOG mRNA水平无明显变化,但NANOG蛋白水平增加,同时细胞增殖能力增强,细胞凋亡水平降低;与空白对照组相比,HAUSP干扰组细胞中,HAUSP的mRNA、蛋白水平表达均降低,NANOG mRNA水平无明显变化,但NANOG蛋白水平降低,同时细胞增殖能力减弱,细胞凋亡水平增加。结论 HAUSP过表达可促进胶质瘤细胞的增殖,这一过程中伴随着NANOG蛋白水平的增高。因此,HAUSP可能通过维持NANOG去泛素化状态而维持胶质瘤细胞的增殖活性。
[Abstract]:Objective to investigate the effects of regulation of herpesvirus-associated ubiquitin specific protease (HAUSP) on the proliferation and apoptosis of glioma cells.Methods overexpression and low expression plasmid of HAUSP were constructed and transfected into U87 U251 glioma cells. The expression level of HAUSP was up-regulated or down-regulated.The expression of HAUSP mRNA and protein in glioma cells was detected by fluorescence quantitative blot, the proliferation of glioma cells was detected by tetramethylazolium colorimetry, and apoptosis was detected by flow cytometry.At the same time, the expression of NANOG mRNA and protein were detected.Results stable and low expression HAUSP overexpression and low expression U87 U251 cell lines were successfully constructed. Compared with the control group, the protein level of HAUSP overexpression group was significantly increased, but the level of NANOG protein was increased.At the same time, the cell proliferation ability was enhanced, and the apoptosis level was decreased. Compared with the blank control group, the expression of mRNAs and protein levels in the HAUSP interference group showed no significant change, but the NANOG protein level was decreased, and the cell proliferation ability was decreased.The level of apoptosis increased.Conclusion overexpression of HAUSP can promote the proliferation of glioma cells, which is accompanied by the increase of NANOG protein level.Therefore, HAUSP may maintain the proliferative activity of glioma cells by maintaining the NANOG deubiquification state.
【作者单位】： 安徽省立医院神经外科脑功能与脑疾病安徽省重点实验室安徽省脑立体定向神经外科研究所;
【基金】：安徽省自然科学基金青年项目(No.1508085QH184)
【分类号】：R739.41
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本文编号：1726597