CD133 glycosylation is enhanced by hypoxia in cultured glioma stem cells
CD133 glycosylation is enhanced by hypoxia in cultured glioma stem cells
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培养的神经胶质瘤干细胞中缺氧会增强 CD133 糖基化
DOI：
10.3892/ijo.2013.1787
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发表时间：
2013-03-01
影响因子：
5.2
通讯作者：
An, Qian
中科院分区：
文献类型：
--
作者：
Lehnus, Kristina S.;Donovan, Laura K.;An, Qian
癌症干细胞（CSC）标志物CD 133在神经胶质瘤中广泛表达，并且主要通过使用结合细胞外糖基化AC 133表位的CD 133/1抗体来使用。然而，CD 133识别可能受到其糖基化模式和氧张力的影响。本研究采用AC 133高表达的多形性胶质母细胞瘤（GBM）细胞系IN 699研究缺氧对CD 133表达和糖基化状态的影响。IN 699细胞在常氧（21%O-2）和低氧（3%O-2）条件下培养。使用糖基化特异性抗体CD 133/1和结合CD 133的未糖基化细胞内残基的ab 19898，通过蛋白质印迹（WB）、qRT-PCR、免疫细胞化学（ICC）和流式细胞术分析CD 133表达。流式细胞仪检测结果显示，ab 19898在常氧和缺氧条件下分别检测到94.1%和96.2%的CD 133（+）细胞。缺氧使CD 133/1表达的CD 133（+）细胞百分比从69%显著增加到92%（p
The cancer stem cell (CSC) marker CD133 is widely expressed in gliomas and employed mostly by use of the CD133/1 antibody which binds the extracellular glycosylated AC133 epitope. CD133 recognition may, however, be affected by its glycosylation pattern and oxygen tension. The present study investigates the effect of oxygen deprivation on CD133 expression and glycosylation status employing a high AC133-expressing glioblastoma multiforme (GBM) cell line, IN699. IN699 cells were cultured under normoxic (21% O-2) and hypoxic (3% O-2) conditions. CD133 expression was analysed by western blotting (WB), qRT-PCR, immunocytochemistry (ICC) and flow cytometry using the glycosylation-specific antibody CD133/1 and ab19898 which binds the unglycosylated intracellular residues of CD133. By flow cytometry, ab19898 detected 94.1% and 96.2% CD133(+) cells under normoxia and hypoxia, respectively. Hypoxia significantly increased the percentage of CD133(+) cells from 69% to 92% using CD133/1 (p