Mojsin M, Topalovic V, Marjanovic Vicentic J, Stevanovic M
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, 11010, Serbia.
Biochemistry (Mosc). 2015 Feb;80(2):202-7. doi: 10.1134/S0006297915020066.
Transcription factor NF-Y belongs to the embryonic stem cell transcription factor circuitry due to its role in the regulation of cell proliferation. We investigated the role of NF-Y in pluripotency maintenance using NT2/D1 cells as one of the best-characterized human embryonal carcinoma cell line. We investigated the efficiency of protein transduction and analyzed the effects of forced expression of short isoform of NF-Y A-subunit (NF-YAs) on NT2/D1 cell growth and expression of SOX2. We found that protein transduction is an efficient method for NF-Y overexpression in NT2/D1 cells. Next, we analyzed the effect of NF-YAs overexpression on NT2/D1 cell viability and detected significant reduction in cell growth. The negative effect of NF-YAs overexpression on NT2/D1 cell pluripotency maintenance was confirmed by the decrease in the level of the pluripotency marker SOX2. Finally, we checked the p53 status and determined that the NF-Y-induced inhibition of NT2/D1 cell growth is p53-independent.
转录因子NF-Y因其在细胞增殖调控中的作用而属于胚胎干细胞转录因子调控网络。我们使用NT2/D1细胞(一种特征明确的人胚胎癌细胞系)研究了NF-Y在多能性维持中的作用。我们研究了蛋白质转导的效率,并分析了NF-Y A亚基短异构体(NF-YAs)的强制表达对NT2/D1细胞生长和SOX2表达的影响。我们发现蛋白质转导是在NT2/D1细胞中过表达NF-Y的有效方法。接下来,我们分析了NF-YAs过表达对NT2/D1细胞活力的影响,检测到细胞生长显著降低。多能性标志物SOX2水平的降低证实了NF-YAs过表达对NT2/D1细胞多能性维持的负面影响。最后,我们检查了p53状态,确定NF-Y诱导的NT2/D1细胞生长抑制与p53无关。