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硒对人多形性胶质母细胞瘤细胞系的体外作用:一项初步研究。

In Vitro Effects of Selenium on Human Glioblastoma Multiforme Cell Lines: A Preliminary Study.

出版信息

Acta Clin Croat. 2017 Mar;56(1):48-57. doi: 10.20471/acc.2017.56.01.08.

Abstract

Glioblastoma multiforme (GBM) is caused by the central nervous system-derived glial cells, and represents the most common (50%-60%) form of primary brain tumors. The aim of this study was to investigate the in vitro effects of selenium on human GBM cells. In the present study, GMS-10 and DBTRG-05MG human GBM cell lines were used as a model to examine selenium entering the cell, cell proliferation, cytotoxicity, DNA fragmentation and Ki-67 protein expression in selenomethionine treated and non-treated groups. Seleno-L-methionine (SeMet) as the organic source of selenium exerted effects on cell proliferation and cytotoxicity, as assessed with WST-1 and lactate dehydrogenase (LDH) tests, respectively. Apoptosis was assessed by DNA fragmentation with an enzyme-linked immunosorbent assay. Ki-67 protein expression was determined by Western blotting, while selenium measurements were performed in the supernatants and lysates by using Graphite Furnace Atomic Absorption Spectrometry. This is the first study to examine the effects of SeMet on cell proliferation and death in GMS-10 and DBTRG-05MG cells. Both GBM cell lines responded to SeMet in a dose- and time-dependent manner. WST-1 test showed that low-dose SeMet treatment (50 and 100 μM) increased cell proliferation. Analysis of intracellular SeMet levels by using AAS showed results consistent with viability and cytotoxicity tests. SeMet treatment for 72h caused increased DNA fragmentation in both cell lines. In conclusion, our results suggest that SeMet induces cell death at high doses, while increasing cell proliferation at low doses. In the view of the data obtained in this investigation, further studies focusing on the possibility of using SeMet against different types of GBM and in combination with prospect synergic compounds are considered to be worthwhile.

摘要

多形性胶质母细胞瘤(GBM)由中枢神经系统衍生的神经胶质细胞引起,是原发性脑肿瘤中最常见的(50%-60%)类型。本研究旨在探讨硒对人GBM细胞的体外作用。在本研究中,GMS-10和DBTRG-05MG人GBM细胞系被用作模型,以检测硒进入细胞的情况、细胞增殖、细胞毒性、DNA片段化以及在经硒代蛋氨酸处理和未处理组中Ki-67蛋白的表达。分别通过WST-1和乳酸脱氢酶(LDH)试验评估,作为硒有机来源的硒代-L-蛋氨酸(SeMet)对细胞增殖和细胞毒性产生影响。通过酶联免疫吸附测定法检测DNA片段化来评估细胞凋亡。通过蛋白质印迹法测定Ki-67蛋白的表达,同时使用石墨炉原子吸收光谱法测定上清液和裂解物中的硒含量。这是第一项研究SeMet对GMS-10和DBTRG-05MG细胞增殖和死亡影响的研究。两种GBM细胞系对SeMet均呈现剂量和时间依赖性反应。WST-1试验表明,低剂量SeMet处理(50和100μM)可增加细胞增殖。使用原子吸收光谱法分析细胞内SeMet水平,结果与活力和细胞毒性试验一致。SeMet处理72小时导致两种细胞系的DNA片段化增加。总之,我们的结果表明,SeMet在高剂量时诱导细胞死亡,而在低剂量时增加细胞增殖。鉴于本研究获得的数据,进一步研究聚焦于使用SeMet对抗不同类型GBM以及与潜在协同化合物联合使用的可能性,被认为是值得的。

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