Role of HFSCARC in hair follicle stem cell proliferation and differentiation in cashmere goat

Niu Wang, Weidong Zhang, Yi Guo, Fan Zhang, Tongtong Zhang, Xin Wang
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Abstract

As a model of regenerative medicine, hair follicle stem cell (HFSC) plays a determining role in the hair cycle. Emerging evidences showed that long noncoding RNAs regulated the biological function of HFSC. In this current study, we found that lncRNA-000552, standing for “goat secondary HFSC Associated SYNE3 Regulator of HF Cycle” (HFSCARC) expressed higher in anagen than that in telogen of cashmere goat. Through experiments involving nucleocytoplasmic separation and RNA-FISH, we determined that HFSCARC was primarily located in the nucleus of HFSC. To understand the function of HFSCARC, the study performed various assays, including crystal violet staining, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, EdU, and flow cytometry analysis, which collectively revealed that HFSCARC inhibited HFSC proliferation. Additionally, HFSCARC promoted the differentiation of HFSC by investigating the expression of marker genes specific to stem cells and keratinocytes. RNA sequencing analysis was conducted to investigate the global gene expression changes associated with HFSCARC expression. The results showed that HFSCARC altered the expression of genes involved in cell proliferation, hair follicle development, and regulation of bone morphogenetic proteins (BMP) signaling. Furthermore, the study revealed that HFSCARC activated the BMP signaling pathway. Intriguingly, the study found a decreased expression of SYNE3, which was a neighboring gene of HFSCARC. The altered expression of genes associated with transmethylase and demethylase further suggested that HFSCARC might play an important role in regulating the SYNE3 expression. Overall, this study provides valuable insights into the regulatory role of HFSCARC in the biological function of HFSC. These findings contribute to a better understanding of the involvement of noncoding RNAs in the regulation of hair cycle.

Abstract Image

HFSCARC 在羊绒山羊毛囊干细胞增殖和分化中的作用
作为再生医学的一种模式,毛囊干细胞(HFSC)在毛发周期中起着决定性作用。新的证据表明,长非编码 RNA 可调控毛囊干细胞的生物学功能。在本研究中,我们发现lncRNA-000552,即 "山羊次级HFSC相关SYNE3 HF周期调节因子"(HFSCARC)在山羊毛发生长期的表达量高于在毛发衰退期的表达量。通过核胞质分离和 RNA-FISH 实验,我们确定 HFSCARC 主要位于 HFSC 的细胞核中。为了解 HFSCARC 的功能,该研究进行了多种实验,包括水晶紫染色、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑、EdU 和流式细胞术分析,这些实验共同揭示了 HFSCARC 可抑制 HFSC 增殖。此外,通过研究干细胞和角质形成细胞特异性标记基因的表达,HFSCARC 还能促进 HFSC 的分化。为了研究与HFSCARC表达相关的全局基因表达变化,我们进行了RNA测序分析。结果显示,HFSCARC改变了细胞增殖、毛囊发育和骨形态发生蛋白(BMP)信号调控相关基因的表达。此外,研究还发现 HFSCARC 激活了 BMP 信号通路。耐人寻味的是,研究发现与 HFSCARC 邻近的 SYNE3 基因表达量减少。与跨甲基化酶和去甲基化酶相关的基因表达的改变进一步表明,HFSCARC 可能在调控 SYNE3 的表达中发挥了重要作用。总之,本研究为了解 HFSCARC 在 HFSC 生物功能中的调控作用提供了有价值的见解。这些发现有助于更好地理解非编码 RNA 在毛发周期调控中的参与作用。
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