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ChIP-Seq analysis reveals PRKACB as a target gene of HOXC13 involved in rabbit hair follicle development

文献类型： 外文期刊

作者： Chen, Yang 1 ; Bao, Zhiyuan 1 ; Yao, Fan 1 ; Liu, Yan 2 ; Zhao, Bohao 1 ; Wu, Xinsheng 1 ;

作者机构： 1.Yangzhou Univ, Coll Anim Sci & Technol, Yangzhou 225009, Jiangsu, Peoples R China

2.Zhejiang Acad Agr Sci, Anim Husb & Vet Res Inst, Hangzhou 310021, Zhejiang, Peoples R China

关键词： HOXC13; ChIP-Seq; PRKACB; Dermal papilla cells; Hair follicle

期刊名称：GENE （ 2022影响因子：3.5； 五年影响因子：3.3 ）

ISSN： 0378-1119

年卷期： 2024 年 893 卷

页码：

收录情况： SCI

摘要： Dermal papilla cells (DPCs) are key regulators of hair follicle (HF) development and growth, which not only regulate HF growth and cycling but play a role in the pathogenesis of hair loss. The transcription factor Homeobox C13 (HOXC13) can modulate the growth and development of HFs. Nevertheless, the specific genes and pathways regulated by HOXC13 in DPCs have yet to be determined. Thus, to gain a better understanding of genomic binding sites involved in HOXC13-regulated HF development, chromatin immunoprecipitation followed by high throughput sequencing (ChIP-Seq) was performed on rabbit DPCs with pcDNA3.1-3 x Flag-HOXC13 overexpression. A complete set of 9670 enrichment peaks was acquired by applying HOXC13-Flag ChIP. Subsequently, the peak sequence was annotated to the rabbit genome, revealing that 6.1 % of the peaks were identified within in the promoter region. Thereafter, five annotated genes were verified using RT-qPCR. The peak-associated genes were mainly enriched in signaling pathways related to HF development, such as MAPK and PI3K-Akt. Furthermore, by using a dual-luciferase reporter assay, we found that HOXC13 can target the protein kinase cAMP-dependent catalytic beta (PRKACB) promoter region (-1596 similar to -1107 bp) and inhibit its transcription, which was consistent with data obtained from ChIP-seq analysis. Overexpression of PRKACB gene significantly modulated the expression of BCL2, WNT2, LEF1, and SFRP2 genes related to HF development as determined by RT-qPCR (P < 0.01, P < 0.05). The CCK-8 and flow cytometry assays showed that PRKACB significantly inhibited the proliferation of DPCs and promoted apoptosis (P < 0.01). In conclusion, our research revealed that PRKACB has the potential to serve as a novel target gene of HOXC13, contributing to the regulation of the proliferation and apoptosis of DPCs. The process of identifying global target genes can contribute to the understanding of the intricate pathways that HOXC13 regulates in the growth of HFs.

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