本文選題：miR--p + 黑色素皮質(zhì)素受體��； 參考：《中國生物化學(xué)與分子生物學(xué)報》2017年06期

【摘要】：黑色素皮質(zhì)素1受體(MC1R)是在黑色素細(xì)胞內(nèi)表達(dá)的G蛋白耦合受體(G protein-coupled receptor,GPCR)家族成員,參與黑色素細(xì)胞中黑色素的生成。微RNAs(miRNAs)是一類非編碼RNA,通過與靶基因3'-UTR結(jié)合抑制基因表達(dá)。已有研究證明,miR-338-3p在多種人類腫瘤細(xì)胞中(過)表達(dá),可通過下調(diào)靶基因表達(dá)抑制腫瘤細(xì)胞的侵襲遷移能力。然而,有關(guān)miR-338-3p對羊駝皮膚黑色素細(xì)胞的黑色素合成影響卻罕見報道。本研究證明,miRNA-338-3p通過靶向抑制MC1R基因表達(dá),抑制羊駝黑色素細(xì)胞黑色素的生成。采用生物信息學(xué)預(yù)測MC1R基因是miRNA-338-3p的靶基因,其基因表達(dá)抑制羊駝黑色素細(xì)胞黑色素合成。隨后構(gòu)建miR-338-3p真核表達(dá)載體。其基因轉(zhuǎn)染結(jié)合q PT-PCR和Western印跡結(jié)果揭示,與對照細(xì)胞比較,過表達(dá)miRNA-338-3p的羊駝黑色素細(xì)胞的MC1R基因,及其下游與黑色素生成相關(guān)的小眼相關(guān)性轉(zhuǎn)錄因子(MITF)、酪氨酸酶(TYR)、酪氨酸酶相關(guān)蛋白1(TYRP1)、酪氨酸酶相關(guān)蛋白2(TYRP2)編碼基因mRNA及蛋白質(zhì)表達(dá)水平明顯下調(diào)。酶聯(lián)免疫吸附分析顯示,過表達(dá)miRNA-338-3p的羊駝皮膚黑色素細(xì)胞的黑色素產(chǎn)量,較對照細(xì)胞顯著下降(P0.01)。綜上結(jié)果,miR-338-3p可通過抑制靶基因MC1R表達(dá),下調(diào)其下游基因MITF、TYR、TYRP1和TYRP2基因的表達(dá),從而抑制羊駝皮膚黑色素細(xì)胞黑色素的合成。miRNA-338-3p在羊駝生長發(fā)育過程中,是否參與調(diào)控體內(nèi)皮膚黑色素細(xì)胞的黑色素生成尚待進(jìn)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.
【作者單位】： 山西農(nóng)業(yè)大學(xué)動物科技學(xué)院;河北農(nóng)業(yè)大學(xué)動物醫(yī)學(xué)院;
【基金】：山西省科技攻關(guān)項(xiàng)目(No.20140311019-3) 山西農(nóng)業(yè)大學(xué)博士科研啟動項(xiàng)目(No.XB2009022)資助~~
【分類號】：S829.9

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