College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, 030801, China.
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, 030801, China.
Acta Histochem. 2020 Apr;122(3):151508. doi: 10.1016/j.acthis.2020.151508. Epub 2020 Jan 21.
Melanoma is a highly invasive malignant skin tumor having high metastatic rate and poor prognosis. The biology of melanoma is controled by miRNAs. The miRNA-183 cluster, which is composed of miRNA-183∼96∼182 genes, plays an important roles in tumor development. In order to investigate the role and action of miRNA-183 cluster in B16 cells, we overexpressed and knocked down miRNA-183 cluster in B16 cells. Using bioinformatics analysis, we predicted that the key framscript factor of melangenic genes. Microphthalmia-associated transcription factor (MITF) is one of the targets of miRNA-183 cluster. The results of Luciferase activity assays confirmed that MITF was targeted by miRNA-183 cluster. Overexpression and knockdown of miRNA-183 cluster in B16 cells resulted in down and up regulation of MITF expression, respectively at both mRNA and protein levels. Furthmore, overexpression and knockdown of the miRNA-183 cluster in B16 cells decreased and increased the expression of mRNA and protein of melangenic genes tyrosinase (TYR), and tyrosinase-related protein 1 (TYRP1), dopachrome-tautomerase (DCT), as well as the production of melanins and eumelanin production, respectively. On the proliferation and migration pathway, overexpression and knockdown of miRNA-183 cluster increased and decreased, respectively the expression of mRNA and protein of mitogen-activated protein kinase 1 (MEK1), extracellular regulated protein kinases1/2 (ERK1/2) and cAMP-responsive-element binding protein (CREB). These results indicated that miRNA-183 cluster regulated melanogenesis in B16 cells as well as cell proliferation and migration by directly targeting MITF through migration pathway.
黑色素瘤是一种具有高侵袭性的恶性皮肤肿瘤，转移率高，预后差。miRNA 调控黑色素瘤的生物学行为。miRNA-183 簇由 miRNA-183∼96∼182 基因组成，在肿瘤发生发展中发挥重要作用。为了研究 miRNA-183 簇在 B16 细胞中的作用和机制，我们在 B16 细胞中转染 miRNA-183 簇过表达和敲低载体。通过生物信息学分析，预测黑色素瘤基因关键转录因子小眼畸形相关转录因子（MITF）是 miRNA-183 簇的靶基因之一。荧光素酶活性实验证实 MITF 是 miRNA-183 簇的靶基因。miRNA-183 簇在 B16 细胞中的过表达和敲低分别导致 MITF 的 mRNA 和蛋白水平下调和上调。此外，miRNA-183 簇在 B16 细胞中的过表达和敲低分别降低和增加黑色素瘤基因酪氨酸酶（TYR）、酪氨酸酶相关蛋白 1（TYRP1）、多巴色素互变异构酶（DCT）以及黑色素和真黑色素产生的 mRNA 和蛋白的表达。在增殖和迁移途径中，miRNA-183 簇的过表达和敲低分别增加和降低丝裂原活化蛋白激酶 1（MEK1）、细胞外调节蛋白激酶 1/2（ERK1/2）和 cAMP 反应元件结合蛋白（CREB）的 mRNA 和蛋白的表达。这些结果表明，miRNA-183 簇通过直接靶向 MITF 调节 B16 细胞的黑色素生成以及细胞增殖和迁移。
