Bradshaw Amy, Wickremesekera Agadha, Brasch Helen D, Chibnall Alice M, Davis Paul F, Tan Swee T, Itinteang Tinte
Gillies McIndoe Research Institute , Wellington , New Zealand.
Gillies McIndoe Research Institute, Wellington, New Zealand; Department of Neurosurgery, Wellington Regional Hospital, Wellington, New Zealand.
Front Surg. 2016 Aug 26;3:48. doi: 10.3389/fsurg.2016.00048. eCollection 2016.
To identify and characterize cancer stem cells (CSC) in glioblastoma multiforme (GBM).
Four-micrometer thick formalin-fixed paraffin-embedded GBM samples from six patients underwent 3,3-diaminobenzidine (DAB) and immunofluorescent (IF) immunohistochemical (IHC) staining for the embryonic stem cell (ESC) markers NANOG, OCT4, SALL4, SOX2, and pSTAT3. IF IHC staining was performed to demonstrate co-expression of these markers with GFAP. The protein expression and the transcriptional activities of the genes encoding NANOG, OCT4, SOX2, SALL4, and STAT3 were investigated using Western blotting (WB) and NanoString gene expression analysis, respectively.
DAB and IF IHC staining demonstrated the presence of a CSC population expressing NANOG, OCT4, SOX2, SALL4, and pSTAT3 with the almost ubiquitous presence of SOX2 and a relatively low abundance of OCT4, within GBM. The expression of NANOG, SOX2 and, pSTAT3 but, not OCT and SALL4, was confirmed by WB. NanoString gene analysis demonstrated transcriptional activation of NANOG, OCT4, SALL4, STAT3, and SOX2 in GBM.
This study demonstrated a population of CSCs within GBM characterized by the expression of the CSC markers NANOG, SALL4, SOX2, pSTAT3 and OCT4 at the protein and mRNA levels. The almost ubiquitous presence of SOX2 and a relatively low abundance of OCT4 would support the putative existence of a stem cell hierarchy within GBM.
鉴定和表征多形性胶质母细胞瘤（GBM）中的癌症干细胞（CSC）。
对6例患者的4微米厚福尔马林固定石蜡包埋的GBM样本进行3,3-二氨基联苯胺（DAB）和免疫荧光（IF）免疫组织化学（IHC）染色，检测胚胎干细胞（ESC）标志物NANOG、OCT4、SALL4、SOX2和pSTAT3。进行IF IHC染色以证明这些标志物与胶质纤维酸性蛋白（GFAP）的共表达。分别使用蛋白质印迹法（WB）和NanoString基因表达分析研究编码NANOG、OCT4、SOX2、SALL4和STAT3的基因的蛋白质表达和转录活性。
DAB和IF IHC染色显示GBM内存在表达NANOG、OCT4、SOX2、SALL4和pSTAT3的CSC群体，其中SOX2几乎普遍存在，OCT4丰度相对较低。WB证实了NANOG、SOX2和pSTAT3的表达，但未证实OCT和SALL4的表达。NanoString基因分析显示GBM中NANOG、OCT4、SALL4、STAT3和SOX2的转录激活。
本研究证明GBM内存在一群CSC，其特征在于在蛋白质和mRNA水平上表达CSC标志物NANOG、SALL4、SOX2、pSTAT3和OCT4。SOX2几乎普遍存在且OCT4丰度相对较低，这支持了GBM内干细胞层级的假定存在。
