Small RNAseq for B16F1 cells exposed to BrdU or L-Tyr

Purpose: to evaluate changes in the expression of microRNAs, which contribute to explain a phenotype of differential pigmentation and reduction of proliferation, in B16F1 melanoma cells exposed to analogue of tymidine BrdU or aminoacid L-Tyr Methods: Small RNAseq evaluation of the expression profiles of miRNAs in B16F1 melanoma cells exposed to BrdU (0.25 μg / mL) and L-Tyr (5 mM) for 72 h, as well as the expression by RT-qPCR of some molecular targets related with melanogenesis, cell cycle and senescence and construction of network models of regulation and co-expression of microRNAs. Results: It was confirmed that stimulation or melanogenic repression with L-Tyr or BrdU, respectively, generates changes in melanin concentration, reduction in proliferation and changes in expression of microRNAs 470-3p, 470-5p, 30d-5p , 129-5p, 148b-3p, 27b-3p and 211-5p, which presented patterns of coordinated co-expression, related to melanogenesis through their putative targets MITF, Tyr and Tyrp1 and control of cell cycle and senescence: Cyclin D1, Cdk2, Cdk4 and p21. Conclusion: This work improves our understanding of the events of melanogenesis, cell cycle control, and senescence mediated by the coordinated expression of miRNAs and helps to develop new approaches to a molecular biology of melanoma that operates in network The extraction of total RNA, used as input material for small-RNAseq, was performed from three independent replicates of B16F1 cells exposed or not to L-Tyr or BrdU