黑色素细胞中过量表达miR-137对TYRP-1和TYRP-2的影响

The Influences of Over-Expressing miR-137 on TYRP-1 and TYRP-2 in Melanocytes

【目的】证明miR-137通过调控MITF而间接影响小鼠黑色素细胞TYRP-1和TYRP-2的表达,从而影响黑色素的生成。【方法】通过细胞转染技术在细胞水平过量表达miR-137,对黑色素细胞中黑色素含量进行了测定,经RT-PCR以及western blot检测转染后细胞TYRP-1和TYRP-2在mRNA和蛋白水平的变化。【结果】黑色素含量明显减少,在mRNA水平,TYRP1显著降低至0.4倍,TYRP-2显著降低至0.6倍。在蛋白水平,TYRP-1显著降低至0.61倍,TYRP-2显著降低至0.73倍。【结论】过量表达miR-137会通过调控MITF而使黑色素细胞TYRP-1和TYRP-2的表达量降低,从而揭示了miR-137通过调节TYRP-1和TYRP-2的表达间接调控了黑色素的生成。

【Objective】Coat color is one of the most important economic characters.There are as many as hundreds of regulation genes involved in the formation process of coat color,and the mechanism（s） have not been completely determined.TYRP-1 and TYRP-2 are involved in regulation and control of melanin synthesis,and they are members of tyrosinase family.MITF is inhibited by miR-137 at the translational level,however,TYRP-1 and TYRP-2 were regulated by MITF at the transcriptional level.In this study,the effect of miR-137 on the production of TYRP-1 and TYRP-2 by regulating MITF was studied.【Method】 Over-expression of miR-137 was conducted through the cell transfection technique and the contents of melanin in melanocytes were extracted,and then the level of TYRP-1 and TYRP-2 was detected using RT-PCR and western blot.【Result】 The results showed that the contents of melanin were significantly reduced.TYRP1 mRNA was significantly reduced to 0.4 times and TYRP-2 mRNA was significantly reduced to 0.6 times.TYRP-1 protein was significantly reduced to 0.61 times and TYRP-2 protein was significantly reduced to 0.73 times.【Conclusion】 Results of the study suggested that miR-137 indirectly regulates the variation of YRP-1 and TYRP-2 by regulating MITF,and the miR-137 may indirectly mediate the alteration of coat color through regulating the expression of TYRP-1and TYRP-2.