Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor
Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor
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DOI：
10.3389/fcell.2020.00157
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发表时间：
2020-03-18
影响因子：
5.5
通讯作者：
Kiyono, Tohru
中科院分区：
文献类型：
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作者：
Fukuda, Tomokazu;Takahashi, Kouhei;Kiyono, Tohru
雄激素性脱发(AGA)是最常见的脱发类型，主要由睾酮对毛乳头细胞(DPC)的生物学效应引起。体外培养的DPC可能是筛选AGA靶分子的有用工具。然而，由于细胞衰老和细胞培养应激，原代DPC不能持续增殖。在本研究中，我们将突变型细胞周期蛋白依赖性激酶4(CDK4)、细胞周期蛋白D1和端粒酶逆转录酶(TERT)导入DPC。我们证实了CDK4和Cyclin D1的蛋白表达，以及TERT的酶活性。此外，我们还发现所建立的细胞系没有细胞衰老。我们还利用重组逆转录病毒引入了雄激素受体基因，以补偿细胞增殖过程中转录抑制的内源性雄激素受体。此外，我们检测到在双氢睾酮处理后，雄激素受体有效地核转位到细胞核中，表明我们引入的受体具有功能。我们建立的细胞系是识别由睾酮激活的下游信号通路的有用工具。
Androgenetic alopecia (AGA) is the most common type of hair loss, and is mainly caused by the biological effects of testosterone on dermal papilla cells (DPCs). In vitro culturing of DPCs might be a useful tool for the screening of target molecule of AGA. However, primary DPCs cannot continuously proliferate owing to cellular senescence and cell culture stress. In this study, we introduced mutant cyclin-dependent kinase 4 (CDK4), Cyclin D1, and telomerase reverse transcriptase (TERT) into DPCs. We confirmed protein expression of CDK4 and Cyclin D1, and enzymatic activity of TERT. Furthermore, we found the established cell line was free from cellular senescence. We also introduced the androgen receptor gene using a recombinant retrovirus, to compensate the transcriptional suppressed endogenous androgen receptor in the process of cell proliferation. Furthermore, we detected the efficient nuclear translocation of androgen receptor into the nucleus after the treatment of dihydrotestosterone, indicating the functionality of our introduced receptor. Our established cell line is a useful tool to identify the downstream signaling pathway, which activated by the testosterone.