中分化唇鳞状细胞癌中三种癌症干细胞亚群的鉴定
The Identification of Three Cancer Stem Cell Subpopulations within Moderately Differentiated Lip Squamous Cell Carcinoma.
作者信息
Ram Rachna, Brasch Helen D, Dunne Jonathan C, Davis Paul F, Tan Swee T, Itinteang Tinte
机构信息
Gillies McIndoe Research Institute , Wellington , New Zealand.
Gillies McIndoe Research Institute, Wellington, New Zealand; Wellington Regional Plastic, Maxillofacial and Burns Unit, Hutt Hospital, Wellington, New Zealand.
出版信息
Front Surg. 2017 Mar 6;4:12. doi: 10.3389/fsurg.2017.00012. eCollection 2017.
AIM
To identify and characterize cancer stem cells (CSCs) in moderately differentiated lip squamous cell carcinoma (MDLSCC).
METHOD
MDLSCC samples underwent 3,3-diaminobenzidine (DAB) immunohistochemical (IHC) staining for squamous cell carcinoma marker EMA, CSC marker CD44 and embryonic stem cell markers NANOG, octamer-binding transcription factor 4 (OCT4), spalt-like transcription factor 4 (SALL4), sex-determining region Y-box 2 (SOX2), and phosphorylated signal transducer and activator of transcription 3 (pSTAT3). Immunofluorescent IHC staining was performed on two MDLSCC samples. Western blotting (WB) was used to confirm the expression of the aforementioned proteins and their transcription activation was investigated using NanoString and RT-qPCR.
RESULTS
IHC staining demonstrated the presence of (1) an EMA/CD44/SALL4/NANOG/pSTAT3/SOX2/OCT4 CSC subpopulation within the tumor nests (TNs); (2) a CD44/SALL4/NANOG/pSTAT3/SOX2/OCT4 CSC subpopulation; and (3) a CD44/SALL4/NANOG/pSTAT3/SOX2/OCT4 CSC subpopulation within the stroma, between the TNs. NanoString and RT-qPCR confirmed the presence of mRNA for CD44, SALL4, STAT3, SOX2, and OCT4, and WB confirmed the presence of NANOG, pSTAT3, SOX2, and OCT4.
CONCLUSION
This study demonstrates three putative CSC subpopulations within MDLSCC.
目的
鉴定和表征中度分化唇鳞状细胞癌（MDLSCC）中的癌症干细胞（CSC）。
方法
MDLSCC样本进行3,3 - 二氨基联苯胺（DAB）免疫组织化学（IHC）染色，检测鳞状细胞癌标志物EMA、CSC标志物CD44以及胚胎干细胞标志物NANOG、八聚体结合转录因子4（OCT4）、类spalt转录因子4（SALL4）、性别决定区Y框蛋白2（SOX2）和磷酸化信号转导及转录激活因子3（pSTAT3）。对两个MDLSCC样本进行免疫荧光IHC染色。采用蛋白质免疫印迹法（WB）确认上述蛋白的表达，并使用NanoString和逆转录定量聚合酶链反应（RT - qPCR）研究其转录激活情况。
结果
IHC染色显示肿瘤巢（TNs）内存在（1）一个EMA/CD44/SALL4/NANOG/pSTAT3/SOX2/OCT4 CSC亚群；（2）一个CD44/SALL4/NANOG/pSTAT3/SOX2/OCT4 CSC亚群；（3）TNs之间基质内的一个CD44/SALL4/NANOG/pSTAT3/SOX2/OCT4 CSC亚群。NanoString和RT - qPCR证实存在CD44、SALL4、STAT3、SOX2和OCT4的mRNA，WB证实存在NANOG、pSTAT3、SOX2和OCT4。
结论
本研究证明MDLSCC内存在三个假定的CSC亚群。
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