miRNA-338-3p通过靶向抑制MC1R基因表达抑制羊驼黑色素细胞黑色素的生成

发布时间：2018-04-29 18:38

本文选题：miR--p + 黑色素皮质素受体　；参考：《中国生物化学与分子生物学报》2017年06期

【摘要】：黑色素皮质素1受体(MC1R)是在黑色素细胞内表达的G蛋白耦合受体(G protein-coupled receptor,GPCR)家族成员,参与黑色素细胞中黑色素的生成。微RNAs(miRNAs)是一类非编码RNA,通过与靶基因3'-UTR结合抑制基因表达。已有研究证明,miR-338-3p在多种人类肿瘤细胞中(过)表达,可通过下调靶基因表达抑制肿瘤细胞的侵袭迁移能力。然而,有关miR-338-3p对羊驼皮肤黑色素细胞的黑色素合成影响却罕见报道。本研究证明,miRNA-338-3p通过靶向抑制MC1R基因表达,抑制羊驼黑色素细胞黑色素的生成。采用生物信息学预测MC1R基因是miRNA-338-3p的靶基因,其基因表达抑制羊驼黑色素细胞黑色素合成。随后构建miR-338-3p真核表达载体。其基因转染结合q PT-PCR和Western印迹结果揭示,与对照细胞比较,过表达miRNA-338-3p的羊驼黑色素细胞的MC1R基因,及其下游与黑色素生成相关的小眼相关性转录因子(MITF)、酪氨酸酶(TYR)、酪氨酸酶相关蛋白1(TYRP1)、酪氨酸酶相关蛋白2(TYRP2)编码基因mRNA及蛋白质表达水平明显下调。酶联免疫吸附分析显示,过表达miRNA-338-3p的羊驼皮肤黑色素细胞的黑色素产量,较对照细胞显著下降(P0.01)。综上结果,miR-338-3p可通过抑制靶基因MC1R表达,下调其下游基因MITF、TYR、TYRP1和TYRP2基因的表达,从而抑制羊驼皮肤黑色素细胞黑色素的合成。miRNA-338-3p在羊驼生长发育过程中,是否参与调控体内皮肤黑色素细胞的黑色素生成尚待进一步研究。
[Abstract]:Melanin 1 receptor (MC1R) is a member of G protein coupled receptor G protein-coupled receptor (GPCR) family expressed in melanocytes, which is involved in the production of melanin in melanocytes. MicroRNAs-miRNAs) is a class of noncoding RNAs that inhibit gene expression by binding to target gene 3'-UTR. It has been shown that the expression of miR-338-3p in various human tumor cells can inhibit the invasion and migration of tumor cells through down-regulation of target gene expression. However, the effect of miR-338-3p on melanin synthesis in alpaca skin melanocytes is rarely reported. This study demonstrated that miRNA-338-3p inhibits melanin production in alpaca melanocytes by targeting MC1R gene expression. Bioinformatics was used to predict that MC1R gene was the target gene of miRNA-338-3p, and its gene expression inhibited melanin synthesis in alpaca melanocytes. Then the eukaryotic expression vector of miR-338-3p was constructed. The results of gene transfection combined with Q PT-PCR and Western blotting revealed that the MC1R gene of ALM cells overexpressed with miRNA-338-3p was higher than that of control cells. The mRNA and protein expression levels of the transcriptional factor MITF, tyrosinase, tyrosinase associated protein 1, tyrosinase associated protein 2 and tyrosinase associated protein 2TYRP2were significantly down-regulated in the lower reaches of MITFN, tyrosinase associated protein 1, and tyrosinase associated protein 2TYRP2. Enzyme-linked immunosorbent assay (Elisa) showed that melanin production of melanocytes in alpaca skin was significantly lower than that of control cells. The results showed that miR-338-3p could inhibit the synthesis of melanocyte melanin. MiRNA-338-3p could inhibit the growth and development of alpaca by inhibiting the expression of target gene MC1R and down-regulating the expression of the downstream genes MITF-TYRRP1 and TYRP2. Whether it is involved in the regulation of melanocyte melanogenesis in vivo remains to be further studied.
【作者单位】：山西农业大学动物科技学院;河北农业大学动物医学院;
【基金】：山西省科技攻关项目(No.20140311019-3) 山西农业大学博士科研启动项目(No.XB2009022)资助~~
【分类号】：S829.9

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