黑色素细胞中过量表达lpa-miR-nov-66对TYRP2的影响  

The Influences of over-expressing lpa-miR-nov-66 on TYRP2 in melanocytes of alpaca

作　　者：杨姗姗[1] 范瑞文[1] 焦丁兴 董常生[1] 

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]河北省清河县农业局,河北清河054000

出　　处：《中国兽医杂志》2017年第10期24-27,共4页Chinese Journal of Veterinary Medicine

基　　金：国家"863"计划(2013AA102506);公益性行业(农业)科研专项(201303119);山西省研究生教育创新项目(2016BY071)

摘　　要：本研究旨在探讨lpa-miR-nov-66对黑色素生成相关基因TYRP2的影响。采用细胞转染技术,在羊驼黑色素细胞中过表达lpa-miR-nov-66,运用反转录聚合酶链反应(RT-PCR)、实时荧光定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)技术和免疫印迹法(western blotting)技术分别检测转染lpa-miR-nov-66后黑色素细胞内TYRP2在mRNA和蛋白水平的表达差异。RT-PCR结果显示:TYRP2的部分片段扩增,扩增片段长度为151 bp。qRT-PCR和免疫印迹(western blotting)结果显示:与对照组相比,实验组TYRP2在mRNA表达水平显著降低,降低0.44倍;蛋白表达水平也显著降低,降低0.78倍。结果表明,lpa-miR-nov-66可以影响TYRP2的表达,可能参与了动物毛色的形成过程。The molecular mechanisms underlying the formation of coat colors in animals are poorly understood. Recent studies have demonstrated that mieroRNAs play important roles in the control of melanogenesis and coat color in mammals. In order to inves- tigate the influences of lpa-miR-nov-66 on TYRP2 in melanocytes. The metanocytes were cultured and transfected with the lpa-miR- nov-66 or NC plasmid. Reverse tran^ription- polymerase chain reaction （RT-PCR） , quantitative real -time polymerse chain reac- tion （qRT-PCR） and western blotting were used to detect the different expression of TYRI~ in melanocytes transfeeted with the lpa- miR-nov-66 or NC plasmid. A partial ,sequence of TYRP2 gene with the length of 151 bp was successfully amplified by RT-PCR. qRT-PCR results showed that the expression level of TYRP2 mRNA was 0.44 fold changes in the melanocytes transfected with lpa- miR-nov - 66 plasmid compared with the melanocytes transfected with NC plasmid （ P 〈 0.001 ）. Western blotting showed that the protein level of TYRP2 was 0.78 fold changes in in the melanocytes transfected with lpa-miR-nov--66 plasmid compared with the mel- anoeytes transfected with NC plasmid （ P 〈 0.01 ）. All those results suggest that lpa-miR-nov-66 can affect the expression of TYRP2 both at mRNA and protein levels and has a potential role in coat color regulation.

关 键 词：mircoRNA lpa-miR-nov-66 黑色素细胞 TYRP2 

分 类 号：S852.1[农业科学—基础兽医学]