microRNA-146a通过靶向MAPK4和Myosin Va基因调控羊驼黑色素细胞增殖、迁移

doi:10.11843/j.issn.0366-6964.2021.04.012

Abstract

Abstract: This study aimed to explore the regulatory role of miR-146a on the proliferation and migration of alpaca melanocytes and its molecular mechanism. The dual luciferase assay was used to verify that MAPK4 and Myosin Va were the target genes of miR-146a by co-transfection with MAPK4 or Myosin Va in 293T cells; Quantitative real-time PCR and Western blotting were used to detect the expression of downstream genes in alpaca melanocytes after overexpressing miR-146a; CCK8 and Transwell were used to detect the proliferation and migration of alpaca melanocytes after overexpressing miR-146a. The results showed that, compared to the negative control, dual luciferase activity extremely significantly decreased by 36% or 30% in 293T cells co-transfected with miR-146a and MAPK4 or Myosin Va, respectively (P<0.001); In alpaca melanocytes, with the overexpression of miR-146a, the mRNA expression of MAPK4 and Myosin Va were extremely significantly down-regulated by 67% and 47%(P<0.001, P<0.01), and the protein expression of MAPK4 and Myosin Va were significantly down-regulated by 38% and 69%(P<0.05, P<0.01), respectively; The expression of genes related to proliferation and migration(CREB, MITF, MLPH, Rab27a) were extremely significantly down-regulated at the transcriptional and translational levels(P<0.01, P<0.001); CCK8 and Transwell results showed that overexpression of miR-146a extremely significantly down-regulated the proliferation and migration ability of alpaca melanocytes (P<0.01). In summary, miR-146a targeting MAPK4 and Myosin Va inhibited the proliferation and migration of alpaca melanocytes through downregulating the expression of MEK1, CREB, MITF, MLPH, Rab27a.

Key words: miR-146a, MAPK4, Myosin Va, proliferation, migration

CLC Number:

S813.1

LIU Xuexian, DU Bin, GUO Xiang, XUE Jixuan, YU Leitao, FAN Ruiwen. microRNA-146a Regulates the Proliferation and Migration of Alpaca Melanocytes by Targeting MAPK4 and Myosin Va Genes[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(4): 967-975.

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microRNA-146a Regulates the Proliferation and Migration of Alpaca Melanocytes by Targeting MAPK4 and Myosin Va Genes

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