Hypoxia-inducible factor 1α regulates chemoresistance of glioma cells in hypoxia environment: A preliminary study

Q4 Medicine

中华神经外科杂志 Pub Date : 2019-09-28 DOI:10.3760/CMA.J.ISSN.1001-2346.2019.09.015

Junwei Wang, Pan Wang, Dewei Zou, Bin Liao, Sheng Gong, Lu Zhao

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引用次数: 0

Abstract

Objective To confirm that hypoxia can induce reverse differentiation of glioma cells or maintain the stemness of glioma cells, leading to chemotherapeutic resistance and to explore its mechanism by in vitro studies. Methods U87 and GL261 cells were considered as glioma cells line in this study. CD133+ CD15+ Nestin+ and CD133-CD15-Nestin- glioma cells were sorted through magnetic activated cell sorting and cultured in normoxia and hypoxia environments. CD133+ CD15+ Nestin+ and CD133-CD15-Nestin- glioma cells were cultured in hypoxia to detect the expressions of CD133, CD15, Nestin and hypoxia-inducible factor (HIF) 1α by Western blot and immunofluorescence assay. Cell apoptosis and half maximal inhibitory concentration (IC50) values were assessed after temozolomide (TMZ) treatment. HIF1α knocked-out glioma cells were cultured in hypoxia and the expressions of CD133, CD15 and Nestin were determined by Western blot. Besides, we compared cells apoptosis rate and IC50 values after TMZ treatment between HIF1α knocked-out glioma cells and control group. Results GL261 CD133+ CD15+ Nestin+ cells showed lower cells apoptosis and higher IC50 value compared with Gl261 CD133-CD15-Nestin- cells after TMZ treatment (both P<0.05). The results of Western blot showed that the expression levels of CD133, CD15, Nestin and HIF1α in CD133+ CD15+ Nestin+ and CD133-CD15-Nestin- U87 cells in hypoxia were higher than those in control group cultured in normoxia, which were barely expressed in the latter (all P<0.05). There were lower cells apoptosis and higher IC50 value after TMZ treatment in hypoxia in CD133+ CD15+ Nestin+ and CD133-CD15-Nestin- U87 and GL261 glioma cells (all P<0.05). The expressions of CD133, CD15 and Nestin were decreased in HIF1α knocked-out glioma cells compared with those in cells without knocking out HIF1α. Higher cells apoptosis rate and lower IC50 value were reported for HIF1α knocked-out glioma cells after TMZ treatment (all P<0.05). Conclusion Hypoxia could induce chemotherapy resistance by maintaining the stemness of glioma stem cells or promoting the reverse differentiation of normally differentiated glioma cells. Key words: Glioma; Cell hypoxia; Hypoxia-inducible factor 1, alpha subunit; Chemoresistance

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缺氧诱导因子1α调节神经胶质瘤细胞在缺氧环境中的化疗耐药性的初步研究

目的通过体外研究证实缺氧可诱导胶质瘤细胞反向分化或维持胶质瘤细胞的干性，导致化疗耐药性，并探讨其机制。方法以U87和GL261细胞为神经胶质瘤细胞系。CD133+CD15+Nestin+和CD133-CD15-Nestin-胶质瘤细胞通过磁激活细胞分选进行分选，并在常氧和缺氧环境中培养。CD133+CD15+Nestin+和CD133-CD15-Nestin-神经胶质瘤细胞在缺氧条件下培养，采用免疫荧光和蛋白质印迹法检测CD133、CD15、Nestin和缺氧诱导因子（HIF）1α的表达。替莫唑胺（TMZ）治疗后评估细胞凋亡和半数最大抑制浓度（IC50）值。缺氧培养HIF1α敲除的胶质瘤细胞，用Western印迹法检测CD133、CD15和Nestin的表达。此外，我们比较了HIF1α敲除的神经胶质瘤细胞和对照组TMZ治疗后的细胞凋亡率和IC50值。结果GL261 CD133+CD15+Nestin+细胞经TMZ处理后细胞凋亡率较低，IC50值较GL261 CD133-CD15-Nestin-细胞高（均P<0.05），缺氧TMZ处理后，CD133+CD15+Nestin+、CD133-CD15-Nestin-U87和GL261胶质瘤细胞凋亡率降低，IC50值升高（均P<0.05）。结论缺氧可通过维持胶质瘤干细胞的干性或促进正常分化的胶质瘤细胞的反向分化来诱导化疗耐药性。关键词：胶质瘤；细胞缺氧；缺氧诱导因子1，α亚基；化疗耐药性

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来源期刊

中华神经外科杂志 Medicine-Surgery

CiteScore

0.10

自引率

0.00%

发文量

10706

期刊介绍： Chinese Journal of Neurosurgery is one of the series of journals organized by the Chinese Medical Association under the supervision of the China Association for Science and Technology. The journal is aimed at neurosurgeons and related researchers, and reports on the leading scientific research results and clinical experience in the field of neurosurgery, as well as the basic theoretical research closely related to neurosurgery.Chinese Journal of Neurosurgery has been included in many famous domestic search organizations, such as China Knowledge Resources Database, China Biomedical Journal Citation Database, Chinese Biomedical Journal Literature Database, China Science Citation Database, China Biomedical Literature Database, China Science and Technology Paper Citation Statistical Analysis Database, and China Science and Technology Journal Full Text Database, Wanfang Data Database of Medical Journals, etc.