摘要
旨在证明miR-324-3p可以通过调控其预测的靶基因MC1R及其下游基因的表达从而对羊驼皮肤黑色素的合成产生影响。本研究在体外培养的羊驼皮肤黑色素细胞中转染miR-324-3p过表达载体,应用qRT-PCR与Western blotting分析比较各试验组中MC1R基因与毛色相关基因小眼畸形相关转录因子(Microphthalmia-associtated transcription factor,Mitf)、酪氨酸酶(Tyrosinase,Tyr)、酪氨酸相关蛋白2(Tyrosinase related protein 2,Tyrp2)的表达差异性,利用酶标仪检测黑色素产量的变化。结果显示:(1)miR-324-3p在棕色与白色羊驼皮肤中均有表达,且在棕色羊驼皮肤中极显著表达(P<0.01),其相对表达量是白色的1.64倍;(2)黑色素细胞被转染了miR-324-3p过表达载体后,处理组靶基因MC1R及其下游调控基因Mitf、Tyr和Tyrp2的表达量与黑色素产量较空白对照组均有下调,且以Mitf基因表达量极显著下调(P<0.01)。综上表明,羊驼皮肤中miR-324-3p可能通过调控MC1R基因的表达,顺势下调MC1R基因下游调控基因Mitf、Tyr与Tyrp2的表达,最终对羊驼皮肤黑色素细胞中黑色素的类型及合成量产生影响。
To investigate the effect of miR-324-3p on regulating melanin production by targeting MC1 R,miR-324-3p was over-expressed by liposome transfection method in alpaca melanocytes in vitro.The expression of MC1 R and hair color genes（Mitf,Tyr and Tyrp2）were detected by quantitative real time PCR and Western blotting and the melanin prodution was analyzed by Microplate Reader.The results showed that miR-324-3p was expressed extremely significant difference in the alpaca skins with 1.64 folds in brown alpaca skins vs white alpaca skins（P 〈0.01）,the gene expression of MC1 R,Mitf,Tyr and Tyrp2 were down-regulated and the melanin production was reduced（especially Mitf（P 〈0.01））.The results suggested that miR-324-3p regulated melanogenesis by targeting MC1 Rgene to down-regulate the expression of the downstream genes including Mitf,Tyr and Tyrp2 in the skin of alpaca.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2016年第7期1381-1388,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
山西省科技攻关项目(20140311019-3)
山西农业大学博士科研启动项目(XB2009022)
