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Journal of Integrative Agriculture
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circKIF27 inhibits melanogenesis and proliferation by targeting miR-129-5p/TGIF2 pathway in goat melanocytes
JI Kai-yuan1,2, ZHAO Yi-wei1,2, YUAN Xin1,2, LIANG Chun-e1,2, ZHANG Xue-qing1,2, TIAN Wen-li1,2, YU Tong1,3, MA Yang-yang1, LING Ying-hui1, ZHANG Yun-hai1,3#

1 Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China

2 Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China

3 Linquan Comprehensive Experimental Station of Anhui Agricultural University, Anhui Agricultural University, Linquan 236400, China

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摘要  动物的被皮和毛发是经济动物十分重要的经济性状,其质量的好坏直接影响动物的经济价值。动物被毛的颜色是由皮肤毛囊中的黑色素细胞产生的黑色素决定,研究黑色素的生成规律不仅对指导动物育种具有重要的意义,同时对解析色素沉积和色素生成相关疾病的发病机理有重要的参考价值。Circular RNA-micro RNA (circRNA-miRNA) 网络广泛参与各种生物过程,但其在色素沉着中的作用尚未被解析。前期研究发现,circKIF27的表达水平在白色和棕色山羊皮肤分离的黑色素细胞中存在显著差异,其具体的功能尚不明确。我们通过原位杂交实验、PCR实验证实,circKIF27具有circRNA的结构特点,表达于黑色素细胞的细胞质。表型研究表明,黑色素细胞中过表达circKIF27可以显著降低黑色素产量(P<0.01),抑制细胞增殖(P<0.0001)。通过生物信息学分析发现circKIF27与miR-129-5p存在靶向结合位点,可能存在靶向吸附关系。通过荧光素酶报告实验qRT-PCR实验证实,circKIF27靶向吸附miR-129-5p抑制其表达MiR-129-5p功能研究发现,黑色素细胞中过表达miR-129-5p以提高黑色素产量(P<0.01),促进细胞增殖(P<0.0001)。通过生物信息学分析发现TGIF2存在两个潜在的miR-129-5p结合位点,通过qRT-PCR实验证实miR-129-5p可以抑制TGIF2表达。为进一步研究TGIF2在黑色素细胞中的功能,通过构建siRNA-TGIF2,转染黑色素细胞降低TGIF2表达水平,黑色素的产量(P<0.01)和细胞增殖(P<0.0001)速度都显著提升。以上结果表明,circKIF27通过靶向miR-129-5p/TGIF2通路抑制山羊皮肤黑色素生成。本研究为circRNA-miRNA网络调控动物黑色素形成的分子机制提供了新的见解,这有助于了解哺乳动物皮肤和被毛色素沉着的复杂过程,并为指导动物育种提供参考

Abstract  Skin and hair pigmentation in animals involve intricate regulatory processes. Circular RNA-microRNA (circRNA-miRNA) networks play vital roles in various biological processes, although their involvement in pigmentation has been underexplored. This study focused on circKIF27 expression, which differs significantly in melanocytes isolated from white and brown Boer coat-colored skin, yet its function remains unclear. Here, we investigated the roles of circKIF27 in melanocytes. In situ hybridization assays demonstrated that circKIF27 is expressed in the cytoplasm of melanocytes. qRT-PCR results revealed differential expression levels of circKIF27 in various tissues of male and female goats. Functional analysis showed that circKIF27 overexpression in melanocytes significantly reduces melanin production (P<0.01) and inhibits cell proliferation (P<0.0001). Bioinformatics analysis identified a putative miR-129-5p binding site on circKIF27, and luciferase reporter assays confirmed their interaction. Overexpression of miR-129-5p in melanocytes enhances melanin production (P<0.01) and promotes cell proliferation (P<0.05). Further analysis revealed that TGIF2 possesses two potential miR-129-5p binding sites, and miR-129-5p overexpression in melanocytes significantly inhibits TGIF2 expression (P<0.0001), suggesting a targeted regulatory relationship between these two molecules. Silencing TGIF2 expression via siRNA-TGIF2 transfection leads to increased melanocyte proliferation (P<0.0001) and increased melanin production (P<0.01). These findings highlight the involvement of the circRNA-miRNA network in pigmentation, offering new insights into the molecular mechanisms underlying pigmentation and guiding animal hair color breeding strategies.
Keywords:  Capra hircus        pigmentation        melanocyte        circRNA        miRNA  
Online: 12 March 2024  
About author:  JI Kai-yuan, E-mail: 18234487253@163.com; ZHAO Yi-wei, Email: 1641263262@qq.com; #Correspondence ZHANG Yun-hai, Email: yunhaizhang@ahau.edu.cn

Cite this article: 

JI Kai-yuan, ZHAO Yi-we, YUAN Xin, LIANG Chun-e, ZHANG Xue-qing, TIAN Wen-li, YU Tong, MA Yang-yang, LING Ying-hui, ZHANG Yun-hai. 2024. circKIF27 inhibits melanogenesis and proliferation by targeting miR-129-5p/TGIF2 pathway in goat melanocytes. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2024.02.008

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