Combinatorial expression of cell cycle regulators is more suitable for immortalization than oncogenic methods in dermal papilla cells.
Combinatorial expression of cell cycle regulators is more suitable for immortalization than oncogenic methods in dermal papilla cells.
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DOI：
10.1016/j.isci.2020.101929
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发表时间：
2021-01-22
期刊：
影响因子：
5.8
通讯作者：
Ishii T
中科院分区：
文献类型：
--
作者：
Fukuda T;Furuya K;Takahashi K;Orimoto A;Sugano E;Tomita H;Kashiwagi S;Kiyono T;Ishii T
永生化细胞是一种重要的研究工具，它使用强大的生长特性进行基因产物的功能研究。永生化的哺乳动物细胞主要使用三种方法建立：表达猴空泡病毒40 T抗原（SV 40方法）;人乳头瘤病毒衍生的癌蛋白E6/E7（E6/E7方法）;或组合表达R24 C突变型细胞周期蛋白依赖性激酶4、细胞周期蛋白D1和端粒酶逆转录酶（K4 DT方法）。然而，目前还不清楚哪种方法是体外模型的最佳选择。在这里，我们比较了由SV 40，E6/E7或K4 DT方法产生的永生化人毛乳头细胞的生物学特性和全基因组表达谱。据我们所知，这是第一项全面比较表达谱以确定维持野生型细胞原始性质的最佳永生化方法的研究。这些数据对于旨在建立新的永生化细胞系的科学家来说是有价值的。用SV 40、E6/E7、K4 DT方法对永生化毛乳头细胞的表达谱进行了分析，野生型与K4 DT的距离较近，E6/E7居中，SV 40与K4 DT的距离最远。与SV 40和E6/E7方法相比，K4 DT方法的表达谱最保守。
The immortalized cell is an essential research tool that uses robust growth properties for the functional investigation of gene products. Immortalized mammalian cells have mainly been established using three methods: expression of simian vacuolating virus 40 T antigen (the SV40 method); human papilloma virus-derived oncoprotein E6/E7 (the E6/E7 method); or combinatorial expression of R24C mutant cyclin-dependent kinase 4, cyclin D1, and telomerase reverse transcriptase (the K4DT method). However, it is unclear as to which method is optimal for an in vitro model. Here, we compared the biological characteristics and genome-wide expression profiles of immortalized human dermal papilla cells generated by the SV40, E6/E7, or K4DT method. To our knowledge, this is the first study to comprehensively compare expression profiles to determine the optimal immortalization method for maintaining the original nature of the wild-type cells. These data would be valuable to scientists aiming to establish new immortalized cell lines. Expression profiles of immortalized dermal papilla cells by the SV40, E6/E7, K4DT method were analyzed The distance from wild type to K4DT was close, E6E7 was middle, SV40 was most distant K4DT method showed the most conservative profile when it compared with SV40 and E6E7 method Cell Biology; Stem Cell Research; Technical Aspects of Cell Biology
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