Evaluación de miRNAs en la línea celular de melanoma B16 inducida a pigmentación diferencial y disminución del crecimiento celular por la L-Tirosina y la 5-bromo-2´-deoxiuridina

dc.contributorGómez Grosso, Luis Alberto
dc.contributorFisiología Celular y Molecular
dc.creatorRivera Escobar, Hernán Mauricio
dc.date.accessioned2021-01-15T20:59:48Z
dc.date.available2021-01-15T20:59:48Z
dc.date.created2021-01-15T20:59:48Z
dc.date.issued2020-12-04
dc.identifierhttps://repositorio.unal.edu.co/handle/unal/78784
dc.description.abstractmiRNAs are small non-coding RNAs associated with post-transcriptional gene regulation in melanoma, a cancer of melanocytes, cells specialized in melanin synthesis. The present work aimed to evaluate the differential expression of miRNAs and to establish potential functional relationships through their targets in the B16F1 melanoma cell line under conditions of decreased cell growth and differential pigmentation induced by L-tyrosine (L-Tyr) and 5-Bromo-2'-deoxyuridine (BrdU) in vitro. A reduction in proliferation and changes in melanin concentration was confirmed in B16F1 cells exposed to 2.5 µg. mL-1 BrdU or 5 mM L-Tyr. Using DEseq2, counts obtained by small RNAseq were analyzed and differential expression of 35 miRNAs in cells exposed to L-Tyr, 22 under-expressed, and 14 over-expressed; and of 33 miRNAs by exposure to BrdU, 11 over-expressed and 21 under-expressed. The bioinformatics analysis facilitated the construction of co-expression and miRNA regulation network models together with associated targets by KEGG functional enrichment, with the control of the cell cycle, senescence, and pigmentation. Expression levels of 211-5p, 129-5p, 148b-3p, 470-5p, 470-3p, 27b-3p and 30d-5p microRNAs and Mitf, Tyr, Tyrp1, Dct, Ccnd1, Cdk4 Cdk2 and p21 mRNAs were confirmed by RT-qPCR. The results obtained, improve our understanding of the potential functional associations between miRNAs and gene sets during melanogenesis, cell cycle control, and senescence and propose new scenarios for the study of melanoma.
dc.description.abstractLos miRNAs son RNAs pequeños no codificantes asociados con la regulación post-transcripcional de genes, en melanoma, un cáncer de melanocitos, células especializadas en la síntesis de melanina. El objetivo del presente trabajo fue evaluar la expresión diferencial de miRNAs y establecer potenciales relaciones funcionales a través de sus dianas en la línea celular de melanoma B16F1 bajo condiciones de disminución del crecimiento celular y pigmentación diferencial inducidas por la L-tirosina (L-Tyr) y la 5-bromo-2´-deoxiuridina (BrdU) in vitro. Se confirmó una reducción en la proliferación y cambios en la concentración de melanina en células B16F1 expuestas a BrdU 2.5 µg.mL-1 o a L-Tyr 5 mM. Usando DEseq2, se analizaron los conteos obtenidos por small RNAseq y se determinó la expresión diferencial de 35 miRNAs en células expuestas a L-Tyr, 22 sub-expresados y 14 sobre-expresados; y de 33 miRNAs por exposición a BrdU, 11 sobre-expresados y 21 sub-expresados. El análisis bioinformático, facilitó la construcción de modelos en red de co-expresión y de regulación de miRNAs junto a dianas asociadas por enriquecimiento funcional KEGG, con el control del ciclo celular, senescencia y pigmentación. Los niveles de expresión de los microRNAs 211-5p, 129-5p, 148b-3p, 470-5p, 470-3p, 27b-3p y 30d-5p y de los mRNAs Mitf, Tyr, Tyrp1, Dct, Ccnd1, Cdk4 Cdk2 y p21, se confirmaron por RT-qPCR. Los resultados obtenidos, mejoran nuestra comprensión de las potenciales asociaciones funcionales entre conjuntos de miRNAs y genes durante la melanogénesis, el control del ciclo celular y la senescencia y propone nuevos escenarios para el estudio del melanoma.
dc.languagespa
dc.publisherBogotá - Medicina - Doctorado en Ciencias Biomédicas
dc.publisherUniversidad Nacional de Colombia - Sede Bogotá
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dc.rightsAtribución-NoComercial-SinDerivadas 4.0 Internacional
dc.rightsAcceso abierto
dc.rightshttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rightsDerechos reservados - Universidad Nacional de Colombia
dc.titleEvaluación de miRNAs en la línea celular de melanoma B16 inducida a pigmentación diferencial y disminución del crecimiento celular por la L-Tirosina y la 5-bromo-2´-deoxiuridina
dc.typeOtro


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