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Dongsheng Yan, Guangying Luo, Yunping Zhao, Dan-Ning Hu; The Role of Cytosine-5 RNA Methylation in Regulating Uveal Melanoma Cell Proliferation and Migration. Invest. Ophthalmol. Vis. Sci. 2019;60(9):745. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
N5-methylcytosine (m5C) RNA methylation is a novel type of epitranscriptomic modifications, which is catalyzed by the RNA methyltransferases NSUN2 and DNMT2. Emerging evidence indicates that m5C RNA modification contributes to regulating diverse biological processes including gene expression. However, its involvement in uveal melanoma tumorigenesis remains unclear. In this study, we investigated the role of m5C RNA methylation in regulating uveal melanoma cell proliferation and migration.
Dot blot determined m5C RNA methylation levels in both uveal melanoma cell lines and uveal melanocytes. Uveal melanoma cells were transfected with siRNA targeting NSUN2 using Lipofectamine RNAiMAX reagent. MTS and transwell assay were carried out to evaluate the subsequent effects on cell proliferation and migration, respectively. Western blot analysis was performed to detect protein expression levels in uveal melanoma cells.
The RNA m5C methylation level was dramatically upregulated in uveal melanoma cells as compared to uveal melanocytes. Knockdown of NSUN2 led to a dramatic decrease of RNA m5C methylation. Furthermore, downregulation of NSUN2 significantly suppressed uveal melanoma cell proliferation and migration. In addition, NSUN2 knockdown resulted in an upregulation of P21 in uveal melanoma cells.
Our results demonstrate that NSUN2-mediated m5C RNA methylation contributes to regulating uveal melanoma cell proliferation and migration. Targeting such an epitranscriptomic modification may be a promising avenue to treat uveal melanoma.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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