Reciprocal Changes in miRNA Expression with Pigmentation and Decreased Proliferation Induced in Mouse B16F1 Melanoma Cells by l-Tyrosine and 5-Bromo-2′-Deoxyuridine

Abstract

Background: Many microRNAs have been identified as critical mediators in the progression of melanoma through its regulation of genes involved in different cellular processes such as melanogenesis, cell cycle control, and senescence. However, microRNAs’ concurrent participation in syngeneic mouse B16F1 melanoma cells simultaneously induced decreased proliferation and differential pigmentation by exposure to 5-Brd-2′-dU (5’Bromo-2-deoxyuridine) and L-Tyr (L-Tyrosine) respectively, is poorly understood. Aim: To evaluate changes in the expression of microRNAs and identify which miRNAs in-network may contribute to the functional bases of phenotypes of differential pigmentation and reduction of proliferation in B16F1 melanoma cells exposed to 5-Brd-2′-dU and L-Tyr. Methods: Small RNAseq evaluation of the expression profiles of miRNAs in B16F1 melanoma cells exposed to 5-Brd-2′-dU (2.5 μg/mL) and L-Tyr (5 mM), as well as the expression by qRT-PCR of some molecular targets related to melanogenesis, cell cycle, and senescence. By bioinformatic analysis, we constructed network models of regulation and co-expression of microRNAs. Results: We confirmed that stimulation or repression of melanogenesis with L-Tyr or 5-Brd-2′-dU, respectively, generated 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 and reciprocal co-expression, related to changes in melanogenesis through their putative targets Mitf, Tyr and Tyrp1, and control of cell cycle and senescence: Cyclin D1, Cdk2, Cdk4, p21, and p27. Conclusions: These findings provide insights into the molecular biology of melanoma of the way miRNAs are coordinated and reciprocal expression that may operate in a network as molecular bases for understanding changes in pigmentation and decreased proliferation induced in B16F1 melanoma cells exposed to L-Tyr and 5-Brd-2′-dU.

Changes in cytoarchitecture and mobility in B16F1 melanoma cells induced by 5-Br-2'-dU coincide with Rock2, miRNAs 138-5p and 455-3p reciprocal expressions

“…This result is consistent with whose variations have recently linked to alterations in mobility in squamous cell cancer of the tongue and renal fibrosis [ 17 , 18 ] through their common molecular target Rock2. Small RNAseq sequencing results, recently obtained by our laboratory [ 16 ] confirmed the differential expression of these two microRNAs, but also of another set of microRNAs with high expression (129-5p, 30d-5p, 22 -3p, 335-5p, 193b-3p, and 144-3p) and to the low (211-5p, 128-3p and 23a-3p), that have Rock2 as a molecular target [ 44 ]. On the other hand, Vim's mRNA expression showed an increase of 3.64 X ( Fig.…”

“…Cancer cells in melanoma reorganize their cellular skeleton favoring migratory and invasive processes [ [45] , [46] , [47] ]. Exposure to the thymidine analog 5-Br-2'-dU generates a senescent phenotype in different cellular models [ 27 , 29 , 30 ], which implies the restructuring of the cellular cytoskeleton and variations in the expression of mRNAs and miRNAs involved in melanogenesis, cell cycle control, senescence in melanoma [ 15 , 16 , 33 , 34 ]. However, the association between structural variations and mobility of B16F1 cells is still unknown, as well as the involvement of ROCK2 and miRNAs mediate regulation would be related to mobility in other tumor models [ [47] , [48] , [49] , [50] , [51] ].…”

“…The thymidine analog, 5-Bromo-2'-deoxyuridine (5-Br-2'-dU), incorporates into the DNA, generates instability, and has been widely used in different types of cancer [ 11 , 12 ]. Although its mechanism of action is unknown, its exposure in B16 mouse melanoma cells induces a senescent phenotype together with changes in their extracellular matrix glycoproteins, cell attachment, adhesion-related glycoproteins to tumor cell-cell interaction and actin expression, as well as a reduction in proliferation, pigmentation, and differential expression of mRNAs and microRNAs (miRNAs) [ [13] , [14] , [15] , [16] ]. The induction of the senescent phenotype by genotoxic stress by 5-Br-2'-dU in B16F1 cells is little known.…”

Role of miRNAs in melanin metabolism: Implications in melanin‐related diseases

“…It is presumed that the targets of the miRNAs mentioned above are MITF, TYR, and TYRP1. 83 The potential inhibitory activity of miR-21 on the mela-…”

UVB irradiation differential regulate miRNAs expression in skin photoaging

“…Recently, researchers have found a relationship between miRNA and the occurrence and progression of some skin diseases, such as miRNAs and skin cancers, pigmentation, inflammatory diseases, autoimmune diseases, wound healing, and photoaging. 10 , 11 , 12 , 13 Accounted for 1%∼5% of the human genome miRNAs have about 30% of protein-coding genes regulating, 14 and its importance to the whole body is obvious. Likewise, the mechanism of skin photoaging mediated by UVB exposure remains unknown further research can determine the mechanical relationship between miRNAs and UVB irradiation, and it will aid in the prevention of such injury as well as give diagnostic biomarkers and novel nucleic acid targets for treatment.…”