关于湖羊真皮乳头细胞中 microR-148a 和 microR-10a 的初步研究。
Preliminary study on microR-148a and microR-10a in dermal papilla cells of Hu sheep.
机构信息
College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, China.
Joint international research laboratory of agriculture and agri - product safety of Ministry of Education of China, Yangzhou University, Yangzhou, 225009, China.
出版信息
BMC Genet. 2019 Aug 27;20(1):70. doi: 10.1186/s12863-019-0770-8.
BACKGROUND
Hu sheep, a unique Chinese breed with high reproductive performance, are also well known for their rare white lambskin in China. The quality of lambskin is affected by hair follicles, and dermal papilla cells are an important component of hair follicles that plays a key role in hair follicle growth and development. This study helps elucidate the effect of miR-148a and miR-10a on hair follicle growth and development.
RESULTS
Based on the results of gene chip and high-throughput sequencing, bone morphogenetic protein 7 (BMP7) was used as a research object. Bioinformatics analysis and the dual-luciferase reporter system indicated that, along with Western blot and quantitative real-time polymerase chain reaction (qRT-PCR) that miR-148a and miR-10a target relationships with BMP7. BMP7 was the target gene both for miR-148a and miR-10a by the dual-luciferase reporter system and Western blot. Hu sheep dermal papilla cells were successfully isolated and purified, and after transfecting miR-148a/miR-10a mimics and inhibitors into dermal papilla cells, a Cell Counting Kit-8 (CCK-8) was used to determine that miR-148a/miR-10a inhibited the proliferation of Hu sheep dermal papilla cells. In addition, after the overexpression of miR-148a, the expression levels of Smad3 (P < 0.05), Smad6 (P < 0.05), Smad4 (P < 0.01), and Smad5 (P < 0.01) were significantly higher than those of the control groups. After the inhibition of miR-148a, the expression levels of Smad3 (P < 0.05), Smad4 (P < 0.05), and TGF-β (P < 0.01) were significantly lower than those of the control groups. After the overexpression of miR-10a, the expression levels of Smad1 (P < 0.01), Smad2 (P < 0.05), Smad4 (P < 0.01), Smad5 (P < 0.01), and TGF-β (P < 0.05) were significantly lower than those of the control groups. After the inhibition of miR-10a, the expression levels of Smad1 (P < 0.01) and Smad2 (P < 0.05) were significantly lower than those of the control groups.
CONCLUSIONS
These results revealed the target relationship between miR-148a, miR-10a and BMP7, and the effect of miR-148a and miR-10a on the proliferation of dermal papilla cells. They will provide the basis for a follow-up study on how miR-148a, and miR-10a mediate BMP7 regulation of hair follicle growth and development.
背景
湖羊是中国特有的一种繁殖性能高的品种，以其罕见的白色羔皮而闻名。羔皮的质量受毛囊影响，真皮乳头细胞是毛囊的重要组成部分，在毛囊的生长和发育中起着关键作用。本研究旨在阐明 miR-148a 和 miR-10a 对毛囊生长和发育的影响。
结果
基于基因芯片和高通量测序的结果，选择骨形态发生蛋白 7（BMP7）作为研究对象。生物信息学分析和双荧光素酶报告系统表明，miR-148a 和 miR-10a 与 BMP7 之间存在靶向关系。Western blot 和实时定量聚合酶链反应（qRT-PCR）结果表明，BMP7 是 miR-148a 和 miR-10a 的靶基因。双荧光素酶报告系统和 Western blot 证实 BMP7 是 miR-148a 和 miR-10a 的靶基因。成功分离和纯化了湖羊真皮乳头细胞，将 miR-148a/miR-10a 模拟物和抑制剂转染到真皮乳头细胞中后，通过细胞计数试剂盒-8（CCK-8）测定 miR-148a/miR-10a 抑制湖羊真皮乳头细胞的增殖。此外，过表达 miR-148a 后，Smad3（P<0.05）、Smad6（P<0.05）、Smad4（P<0.01）和 Smad5（P<0.01）的表达水平明显高于对照组。抑制 miR-148a 后，Smad3（P<0.05）、Smad4（P<0.05）和 TGF-β（P<0.01）的表达水平明显低于对照组。过表达 miR-10a 后，Smad1（P<0.01）、Smad2（P<0.05）、Smad4（P<0.01）、Smad5（P<0.01）和 TGF-β（P<0.05）的表达水平明显低于对照组。抑制 miR-10a 后，Smad1（P<0.01）和 Smad2（P<0.05）的表达水平明显低于对照组。
结论
这些结果揭示了 miR-148a、miR-10a 与 BMP7 之间的靶关系，以及 miR-148a 和 miR-10a 对真皮乳头细胞增殖的影响。这些结果为进一步研究 miR-148a、miR-10a 如何介导 BMP7 调节毛囊的生长和发育提供了基础。
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