Koh Sabrina P, Brasch Helen D, de Jongh Jennifer, Itinteang Tinte, Tan Swee T
Gillies McIndoe Research Institute, New Zealand.
Wellington Regional Plastic, Maxillofacial & Burns Unit, Hutt Hospital, Wellington, New Zealand.
Heliyon. 2019 Aug 16;5(8):e02257. doi: 10.1016/j.heliyon.2019.e02257. eCollection 2019 Aug.
Cancer stem cells (CSC), the putative origin of cancer, account for local recurrence and metastasis. We aimed to identify and characterize CSCs within moderately differentiated head and neck cutaneous squamous cell carcinoma (MDHNCSCC). Formalin-fixed paraffin-embedded MDHNCSCC sections of ten patients underwent 3,3-diaminobenzidine (DAB) immunohistochemical (IHC) staining for induced pluripotent stem cell (iPSC) markers OCT4, NANOG, SOX2, KLF4 and c-MYC. Localization of these markers was investigated using immunofluorescence (IF) IHC staining of three of these MDHNCSCC samples. mRNA expression of these iPSC markers in the MDHNCSCC tissue samples was determined by colorimetric hybridization (CISH, n = 6), and reverse-transcription quantitative polymerase chain reaction (RT-qPCR, n = 4). RT-qPCR was also performed on four MDHNCSCC-derived primary cell lines. DAB IHC staining demonstrated expression of all five iPSC markers within all ten MDHNCSCC tissues samples. CISH and RT-qPCR confirmed mRNA expression of all five iPSC markers within all MDHNCSCC tissues samples examined. RT-PCR demonstrated mRNA transcripts of all five iPSC markers in all four MDHNCSCC-derived primary cell lines. IF IHC staining showed co-expression of OCT4 with SOX2 and KLF4 throughout the tumor nests (TNs) and peri-tumoral stroma (PTS). There was an OCT4/NANOG subpopulation within the TNs, and an OCT4/NANOG subpopulation and an OCT4/NANOG subpopulation within the PTS. All iPSC markers were expressed by the endothelium of microvessels within the PTS. Our findings suggest the presence of an OCT4/NANOG/SOX2/KLF4/c-MYC CSC subpopulation within the TNs, PTS and endothelium of microvessels within the PTS; and an OCT4/NANOG/SOX2/KLF4/c-MYC subpopulation exclusively within the PTS in MDHNCSCC. These CSC subpopulations could be a potential novel therapeutic target for treatment of MDHNCSCC.
癌症干细胞（CSC）被认为是癌症的起源，与局部复发和转移有关。我们旨在鉴定和表征中度分化的头颈部皮肤鳞状细胞癌（MDHNCSCC）中的癌症干细胞。对10例患者的福尔马林固定石蜡包埋的MDHNCSCC切片进行3,3-二氨基联苯胺（DAB）免疫组织化学（IHC）染色，检测诱导多能干细胞（iPSC）标志物OCT4、NANOG、SOX2、KLF4和c-MYC。使用其中3个MDHNCSCC样本的免疫荧光（IF）IHC染色研究这些标志物的定位。通过比色杂交（CISH，n = 6）和逆转录定量聚合酶链反应（RT-qPCR，n = 4）测定MDHNCSCC组织样本中这些iPSC标志物的mRNA表达。还对4个MDHNCSCC来源的原代细胞系进行了RT-qPCR。DAB IHC染色显示所有10个MDHNCSCC组织样本中均表达所有5种iPSC标志物。CISH和RT-qPCR证实了在所检测的所有MDHNCSCC组织样本中均有所有5种iPSC标志物的mRNA表达。RT-PCR显示所有4个MDHNCSCC来源的原代细胞系中均有所有5种iPSC标志物的mRNA转录本。IF IHC染色显示OCT4与SOX2和KLF4在整个肿瘤巢（TN）和肿瘤周围基质（PTS）中共表达。在TN中有一个OCT4/NANOG亚群，在PTS中有一个OCT4/NANOG亚群和一个OCT4/NANOG亚群。所有iPSC标志物均由PTS内微血管的内皮细胞表达。我们的研究结果表明，在TN、PTS以及PTS内微血管的内皮细胞中存在OCT4/NANOG/SOX2/KLF4/c-MYC癌症干细胞亚群；在MDHNCSCC中，仅在PTS内存在一个OCT4/NANOG/SOX2/KLF4/c-MYC亚群。这些癌症干细胞亚群可能是治疗MDHNCSCC的潜在新型治疗靶点。
