Quantitative Phosphoproteomic Analysis of the STAT3/IL-6/HIF1α Signaling Network: An Initial Study in GSC11 Glioblastoma Stem Cells
Quantitative Phosphoproteomic Analysis of the STAT3/IL-6/HIF1α Signaling Network: An Initial Study in GSC11 Glioblastoma Stem Cells
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DOI：
10.1021/pr9007927
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发表时间：
2010-01-01
影响因子：
4.4
通讯作者：
Conrad, Charles A.
中科院分区：
文献类型：
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作者：
Nilsson, Carol L.;Dillon, Roslyn;Conrad, Charles A.
包括胶质母细胞瘤（GBM）在内的几种癌症的发生和维持可能是由一小部分称为癌症干细胞（CSCs）的细胞驱动的。干细胞可以提供肿瘤细胞群中的细胞库，这些细胞对用于治疗肿瘤的化疗药物具有难治性。STAT3是与多种干细胞类型调控相关的关键转录因子。最近，一种新的自分泌环（IL-6/STAT3/HIF1 α）在多种肿瘤类型（胰腺、前列腺、肺和结肠）中被观察到。本研究的目的是通过使用JAK2/STAT3磷酸化抑制剂（WP1193）、IL-6刺激和缺氧，探测来自人类肿瘤的胶质母细胞瘤癌症干细胞系（GSC11）中该环的干扰。采用定量磷蛋白组学方法，在高分辨率仪器中使用磷蛋白富集、等压标签化学标记、磷酸肽富集和串联质谱。本研究共鉴定出3414个蛋白。快速免疫印迹技术(
Initiation and maintenance of several cancers including glioblastoma (GBM) may be driven by a small subset of cells called cancer stem cells (CSCs). CSCs may provide a repository of cells in tumor cell populations that are refractory to chemotherapeutic agents developed for the treatment of tumors. STAT3 is a key transcription factor associated with regulation of multiple stem cell types. Recently, a novel autocrine loop (IL-6/STAT3/HIF1 alpha) has been observed in multiple tumor types (pancreatic, prostate, lung, and colon). The objective of this study was to probe perturbations of this loop in a glioblastoma cancer stem cell line (GSC11) derived from a human tumor by use of a JAK2/STAT3 phosphorylation inhibitor (WP1193), IL-6 stimulation, and hypoxia. A quantitative phosphoproteomic approach that employed phosphoprotein enrichment, chemical tagging with isobaric tags, phosphopeptide enrichment, and tandem mass spectrometry in a high-resolution instrument was applied. A total of 3414 proteins were identified in this study. A rapid Western blotting technique (