Lpa-miR-nov-66拮抗IGF1促羊驼黑色素细胞黑色素生成及细胞迁移作用

Lpa-miR-nov-66 Antagonizes IGF1-promoted Melanogenesis and Cell Migration in Alpaca Melanocytes

胰岛素样生长因子1(IGF1)能够促进细胞迁移。我们最近的研究发现,一种新发现的miRNA(lpa-miR-nov-66)可通过靶向抑制可溶性腺苷酸环化酶(soluble guanylate cyclase,s GC)抑制黑色素细胞产生黑色素。然而,当二者联合作用于黑色素细胞时,究竟如何影响黑色素细胞的黑色素产生及细胞迁移尚未见报道。本研究证明,lpa-miR-nov-66可拮抗IGF1的促黑色素细胞的黑色素生成及促细胞迁移作用。ELISA法检测结果揭示,与单纯IGF1处理比较,过表达lpa-miR-nov-66或过表达lpa-miR-nov-66联合IGF1处理可明显降低IGF1诱导羊驼黑色素细胞的c AMP生成水平。实时定量PCR和Western印迹证明,与单纯IGF1处理比较,过表达lpa-miR-nov-66或联合处理可明显降低毛色生成相关基因——MITF、TYR、和TYRP1在黑色素细胞的表达。此外,细胞增殖和细胞划痕实验显示,与单纯IGF1处理比较,过表达lpa-miR-nov-66或联合处理可显著抑制黑色素细胞的迁移。上述结果表明,lpa-miR-nov-66可以减少c AMP产生,抑制IGF1的促黑色素细胞产生黑色素的作用,并抑制IGF1对黑色素细胞增殖和迁移的促进作用。

Insulin-like growth factor 1（ IGF1） promotes cell migration. Our previous study demonstrated that a new miRNA（ named lpa-miR-nov-66） can inhibit melanogenesis by directly targeting soluble guanylate cyclase（ s GC）. However,the effects of lpa-miR-nov-66 in IGF1-mediated melanogenesis and cell migration have not been reported. Here, the results demonstrated that lpa-miR-nov-66 could antagonize the effects of IGF1 on promoting melanogenesis and cell migration. The result of ELISA showed that c AMP production was decreased in melanocytes treated by lpa-miR-nov-66 or combination of IGF1 and lpa-miR-nov-66,in comparison with that in melanocytes treated only by IGF1. The results of real-time PCR and Western blotting showed that the expression levels of microphthalmia transcription factor（ MITF）,tyrosinase（ TYR） and tyrosinase related protein 1（ TYRP1） genes and proteins inmelanocytes were decreased in melanocytes treated by lpa-miR-nov-66 or combination of IGF1 and lpamiR-nov-66,in comparison with that in melanocytes treated only by IGF1. The result of wound scratch assay showed that the cell migration of melanocytes treated by combination of IGF1 and lpa-miR-nov-66 was inhibited compared with that in melanocytes treated only by IGF1. The above results suggested that lpa-miR-nov-66 antagonized the effects of IGF1 on melanogenesis of alpaca melanocytes by reducing c AMP production,therefore to inhibit the melanogenesis,cell migration and proliferation.