Preliminary study on microR-148a and microR-10a in dermal papilla cells of Hu sheep
Preliminary study on microR-148a and microR-10a in dermal papilla cells of Hu sheep
复制标题
湖羊真皮乳头细胞microR-148a和microR-10a的初步研究
DOI：
10.1186/s12863-019-0770-8
复制
发表时间：
2019-08-27
期刊：
影响因子：
2.9
通讯作者：
Sun, Wei
中科院分区：
文献类型：
--
作者：
Lv, Xiaoyang;Gao, Wen;Sun, Wei
背景
湖羊是中国特有的繁殖性能高的品种，在中国也因其珍稀的白色羔皮而闻名。羔皮质量受毛囊影响，而毛乳头细胞是毛囊的重要组成部分，在毛囊生长发育中起关键作用。本研究有助于阐明miR - 148a和miR - 10a对毛囊生长发育的影响。
结果
基于基因芯片和高通量测序结果，以骨形态发生蛋白7（BMP7）为研究对象。生物信息学分析和双荧光素酶报告系统表明，结合蛋白质印迹法和实时定量聚合酶链反应（qRT - PCR），miR - 148a和miR - 10a与BMP7存在靶向关系。通过双荧光素酶报告系统和蛋白质印迹法确定BMP7是miR - 148a和miR - 10a的靶基因。湖羊毛乳头细胞成功分离和纯化，将miR - 148a/miR - 10a模拟物和抑制剂转染到毛乳头细胞后，使用细胞计数试剂盒 - 8（CCK - 8）测定发现miR - 148a/miR - 10a抑制湖羊毛乳头细胞的增殖。此外，miR - 148a过表达后，Smad3（P < 0.05）、Smad6（P < 0.05）、Smad4（P < 0.01）和Smad5（P < 0.01）的表达水平显著高于对照组。miR - 148a抑制后，Smad3（P < 0.05）、Smad4（P < 0.05）和转化生长因子 - β（TGF - β）（P < 0.01）的表达水平显著低于对照组。miR - 10a过表达后，Smad1（P < 0.01）、Smad2（P < 0.05）、Smad4（P < 0.01）、Smad5（P < 0.01）和TGF - β（P < 0.05）的表达水平显著低于对照组。miR - 10a抑制后，Smad1（P < 0.01）和Smad2（P < 0.05）的表达水平显著低于对照组。
结论
这些结果揭示了miR - 148a、miR - 10a与BMP7之间的靶向关系，以及miR - 148a和miR - 10a对毛乳头细胞增殖的影响。它们将为后续研究miR - 148a和miR - 10a如何介导BMP7对毛囊生长发育的调控提供依据。
BackgroundHu 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.ResultsBased 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 withBMP7.BMP7was 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 ofSmad3(P< 0.05), Smad6(P< 0.05),Smad4(P< 0.01), andSmad5(P< 0.01) were significantly higher than those of the control groups. After the inhibition of miR-148a, the expression levels ofSmad3(P< 0.05),Smad4(P< 0.05), andTGF-β(P< 0.01) were significantly lower than those of the control groups. After the overexpression of miR-10a, the expression levels ofSmad1(P< 0.01), Smad2(P< 0.05),Smad4(P< 0.01),Smad5(P< 0.01), andTGF-β(P< 0.05) were significantly lower than those of the control groups. After the inhibition of miR-10a, the expression levels ofSmad1(P< 0.01) andSmad2(P< 0.05) were significantly lower than those of the control groups.ConclusionsThese results revealed the target relationship between miR-148a, miR-10a andBMP7, 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 mediateBMP7regulation of hair follicle growth and development.