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Overexpression of microRNA-130a represses uveal melanoma cell migration and invasion through inactivation of the Wnt/β-catenin signaling pathway by downregulating USP6

Abstract

Uveal melanoma (UM) is a neoplasm arising from melanocytes of the ciliary body, choroid, and iris of the eye, which is the most common primary malignant intraocular tumor. microRNA-130a (miR-130a) has been confirmed to be underexpressed in many types of cancers. Here we aimed to investigate the mechanism whereby miR-130a affects the Wnt/β-catenin signaling pathway by targeting ubiquitin-specific protease 6 (USP6) in UM. Ocular specimens of 62 patients with UM and 42 participants subjected to enucleation due to trauma were collected. In the normal uveal tissues and those from metastatic and non-metastatic UM, we evaluated miR-130a expression by RT-qPCR and then measured mRNA and protein expression of recombinant human mothers against decapentaplegic homolog 4 (SMAD4), USP6, related factors of the Wnt/β-catenin signaling pathway, and epidermal growth factor receptor (EGFR) by RT-qPCR and western blot analysis. Subsequently, the interaction between miR-130a and USP6 was identified by bioinformatics analysis and dual-luciferase reporter gene assay. Next, UM cell migration and invasion abilities, as well as tumor growth in nude mice, were measured through gain- and loss-of-function studies of miR-130a and USP6. miR-130a expression was downregulated in uveal tissues from patients with UM, especially in metastatic uveal tissues. The overall survival of UM patients with low miR-130a expression was shorter than those with high miR-130a expression. USP6 was a target of miR-130a and the overexpression of miR-130a or inhibition of USP6 in UM MUM-2B and MUM-2C cell lines inhibited the expression of Wnt, β-catenin, and EGFR, and activated SMAD4 expression, while reducing UM cell migration and invasion abilities in vitro. The above changes could be reversed by overexpressing USP6 in vitro, whereas overexpressed miR-130a could inhibit the tumor growth in nude mice. Taken together, overexpressed miR-130a inhibited USP6 expression to repress UM cell migration and invasion abilities through inactivating the Wnt/β-catenin signaling pathway, which could be a potential candidate for treatment of UM.

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Fig. 1: Bioinformatics analysis of the differentially expressed genes and their molecular interactions in UM.
Fig. 2: High miR-130a expression predicts a good prognosis for survival of UM patients.
Fig. 3: Upregulated miR-130a reduces cell migration and invasion in UM MUM-2B and MUM-2C cell lines.
Fig. 4: USP6 is a target of miR-130a.
Fig. 5: Silencing USP6 impedes cell migration and invasion via inactivation of the Wnt/β-catenin signaling pathway in UM MUM-2B and MUM-2C cell lines.
Fig. 6: USP6 mediates the influence of miR-130a on UM MUM-2B and MUM-2C cell migration and invasion abilities.
Fig. 7: miR-130 overexpression delays tumor growth of nude mice.

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Correspondence to Chao Zhang.

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Wu, S., Han, M. & Zhang, C. Overexpression of microRNA-130a represses uveal melanoma cell migration and invasion through inactivation of the Wnt/β-catenin signaling pathway by downregulating USP6. Cancer Gene Ther (2021). https://doi.org/10.1038/s41417-021-00377-7

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