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Analysis of Human Stem Cell Transcription Factors.
Cellular Reprogramming ( IF 1.6 ) Pub Date : 2019-07-12 , DOI: 10.1089/cell.2019.0005
Smarakan Sneha 1 , Rohit P Nagare 1 , Pacharia Manasa 1 , Sekar Vasudevan 1 , Aboo Shabna 1 , Trivadi Sundaram Ganesan 1
Affiliation  

Transcription factors NANOG, OCT4, SOX2, and NESTIN are expressed in both human embryonic stem cells (hESCs) and cancer stem cells and they play a crucial role in maintaining characteristics of stemness such as self-renewal and pluripotency. This article evaluates the expression of variants of the main stem cell-specific transcription factors NANOG and OCT4 critically and accurately with specific primers designed for identifying the most important variants that maintain stemness. We have examined four variants of NANOG along with a processed pseudogene and seven variants of OCT4 in human teratocarcinoma cell lines (NTERA2D1, SuSa, GCT-27, and 833KE), hESCs, and ovarian cancer cells by reverse transcriptase-polymerase chain reaction. In addition, we have examined their expression in NTERA2D1 cells on differentiation with all-trans-retinoic-acid. We show that NANOG1 is expressed in all teratocarcinoma cells and can be distinguished from NANOGP8, which is an expressed pseudogene. NANOG2 was not expressed in any of the cell lines, including ESCs. OCT4A was expressed in all cells, whereas the variant OCT4B-variant 3 was expressed only in NTERA2D1 cells. On differentiation of NTERA2D1 with retinoic acid, only NANOGP8 and OCT4A were expressed. In ovarian cancer cells, only 3/6 expressed NANOG1 and OCT4A. All malignant cells from patients with ovarian cancer (N = 6) expressed NANOG1 and OCT4A. These results demonstrate the necessity to precisely evaluate the expression of stem cell transcription factors when defining stemness.

中文翻译:

人类干细胞转录因子分析。

转录因子NANOG,OCT4,SOX2和NESTIN在人类胚胎干细胞(hESCs)和癌症干细胞中均表达,它们在维持干细胞的特性(如自我更新和多能性)中起着至关重要的作用。本文使用专为鉴定维持干性的最重要变异而设计的特异性引物,准确而准确地评估了主要干细胞特异性转录因子NANOG和OCT4变异的表达。我们已经通过逆转录酶-聚合酶链反应检测了人畸胎癌细胞系(NTERA2D1,SuSa,GCT-27和833KE),hESCs和卵巢癌细胞中NANOG的四个变异体以及经过处理的假基因和OCT4的七个变异体。另外,我们已经检查了它们在全反式视黄酸分化后在NTERA2D1细胞中的表达。我们显示NANOG1在所有畸胎癌细胞中表达,并且可以与NANOGP8区别开,后者是一种表达的假基因。NANOG2在包括ESC在内的任何细胞系中均未表达。OCT4A在所有细胞中表达,而变异OCT4B-variant 3仅在NTERA2D1细胞中表达。在用视黄酸区分NTERA2D1时,仅表达了NANOGP8和OCT4A。在卵巢癌细胞中,只有3/6表达NANOG1和OCT4A。来自卵巢癌(N = 6)患者的所有恶性细胞均表达NANOG1和OCT4A。这些结果表明在定义干性时精确评估干细胞转录因子表达的必要性。NANOG2在包括ESC在内的任何细胞系中均未表达。OCT4A在所有细胞中表达,而变异OCT4B-variant 3仅在NTERA2D1细胞中表达。在用视黄酸区分NTERA2D1时,仅表达了NANOGP8和OCT4A。在卵巢癌细胞中,只有3/6表达NANOG1和OCT4A。来自卵巢癌(N = 6)患者的所有恶性细胞均表达NANOG1和OCT4A。这些结果表明在定义干性时精确评估干细胞转录因子表达的必要性。NANOG2在包括ESC在内的任何细胞系中均未表达。OCT4A在所有细胞中表达,而变异OCT4B-variant 3仅在NTERA2D1细胞中表达。在用视黄酸区分NTERA2D1时,仅表达了NANOGP8和OCT4A。在卵巢癌细胞中,只有3/6表达NANOG1和OCT4A。来自卵巢癌(N = 6)患者的所有恶性细胞均表达NANOG1和OCT4A。这些结果表明在定义干性时精确评估干细胞转录因子表达的必要性。来自卵巢癌(N = 6)患者的所有恶性细胞均表达NANOG1和OCT4A。这些结果证明了在定义干性时精确评估干细胞转录因子表达的必要性。来自卵巢癌(N = 6)患者的所有恶性细胞均表达NANOG1和OCT4A。这些结果证明了在定义干性时精确评估干细胞转录因子表达的必要性。
更新日期:2019-11-01
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