本文選題：神經(jīng)膠質(zhì)瘤　切入點：HAUSP　出處：《華中科技大學(xué)學(xué)報(醫(yī)學(xué)版)》2017年06期

【摘要】：目的探討調(diào)控皰疹病毒相關(guān)泛素特異性蛋白酶(HAUSP)對膠質(zhì)瘤細胞增殖和凋亡的影響。方法構(gòu)建HAUSP過表達及低表達質(zhì)粒,轉(zhuǎn)染膠質(zhì)瘤U87、U251細胞,上調(diào)或下調(diào)膠質(zhì)瘤細胞中HAUSP表達水平。熒光定量PCR、Western blot檢測膠質(zhì)瘤細胞中HAUSP mRNA和蛋白表達水平;利用四甲基偶氮唑鹽比色法(MTT)檢測細胞增殖水平;流式細胞術(shù)檢測細胞凋亡變化。同時檢測各組細胞NANOG mRNA和蛋白表達情況。結(jié)果成功構(gòu)建穩(wěn)定HAUSP過表達和低表達U87、U251細胞株;與空白對照組相比,HAUSP過表達組細胞中,HAUSP的mRNA、蛋白水平均明顯增加,NANOG mRNA水平無明顯變化,但NANOG蛋白水平增加,同時細胞增殖能力增強,細胞凋亡水平降低;與空白對照組相比,HAUSP干擾組細胞中,HAUSP的mRNA、蛋白水平表達均降低,NANOG mRNA水平無明顯變化,但NANOG蛋白水平降低,同時細胞增殖能力減弱,細胞凋亡水平增加。結(jié)論 HAUSP過表達可促進膠質(zhì)瘤細胞的增殖,這一過程中伴隨著NANOG蛋白水平的增高。因此,HAUSP可能通過維持NANOG去泛素化狀態(tài)而維持膠質(zhì)瘤細胞的增殖活性。
[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.
【作者單位】： 安徽省立醫(yī)院神經(jīng)外科腦功能與腦疾病安徽省重點實驗室安徽省腦立體定向神經(jīng)外科研究所;
【基金】：安徽省自然科學(xué)基金青年項目(No.1508085QH184)
【分類號】：R739.41
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本文編號：1726597