Transcriptome-wide profiling for melanocytes derived from newborn and adult human epidermis with enhanced proliferation

IF 4.3 3区 材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC ACS Applied Electronic Materials Pub Date : 2024-07-03 DOI:10.1002/cbin.12214
Ai Orimoto, Sayo Kashiwagi, Ayaka Funakoshi, Takashi Shimizu, Tsuyoshi Ishii, Tohru Kiyono, Tomokazu Fukuda
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Abstract

The senescence-associated protein p16INK4A acts as a limiter element in cell-cycle progression. The loss of p16INK4A function is causally related to cellular immortalization. The increase in p16INK4A levels with advancing age was demonstrated in melanocytes. However, the characteristic difference between young and senescent melanocytes affecting immortalization of melanocytes remains unclear. In this study, we generated 10 different cell lines in total from newborn (NB) and adult (AD) primary normal human epidermal melanocytes (NHEM) using four different methods, transduction of CDK4R24C and cyclin D1 (K4D), K4D with TERT (K4DT), SV40 T-antigen (SV40T), and HPV16 E6 and E7 (E6/E7) and performed whole transcriptome sequencing analysis (RNA-Seq) to elucidate the differences of genome-wide expression profiles among cell lines. The analysis data revealed distinct differences in expression pattern between cell lines from NB and AD although no distinct biological differences were detected in analyses such as comparison of cell morphology, evaluation of cell proliferation, and cell cycle profiles. This study may provide useful in vitro models to benefit the understanding of skin-related diseases.

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从新生儿和成人表皮中提取的黑素细胞的全转录组图谱,其增殖能力增强。
衰老相关蛋白p16INK4A是细胞周期进展的限制因子。p16INK4A 功能的丧失与细胞永生化有因果关系。黑色素细胞中的 p16INK4A 水平随着年龄的增长而增加。然而,年轻黑色素细胞和衰老黑色素细胞之间的特征性差异对黑色素细胞永生化的影响仍不清楚。在这项研究中,我们采用了四种不同的方法,即转导 CDK4R24C 和细胞周期蛋白 D1(K4D),从新生(NB)和成年(AD)的原代正常人表皮黑色素细胞(NHEM)中总共生成了 10 种不同的细胞系、K4D with TERT (K4DT)、SV40 T-抗原 (SV40T) 和 HPV16 E6 和 E7 (E6/E7),并进行了全转录组测序分析(RNA-Seq),以阐明细胞系之间全基因组表达谱的差异。分析数据显示,尽管在细胞形态比较、细胞增殖评估和细胞周期图谱等分析中未发现明显的生物学差异,但NB和AD细胞系之间的表达模式存在明显差异。这项研究可提供有用的体外模型,有助于了解皮肤相关疾病。
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来源期刊
CiteScore
7.20
自引率
4.30%
发文量
567
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