Melanogenesis stimulation in B16-F10 melanoma cells induces cell cycle alterations, increased ROS levels and a differential expression of proteins as revealed by proteomic analysis.
Considering that stimulation of melanogenesis may lead to alterations of cellular responses, besides melanin production, our main goal was to study the cellular effects of melanogenesis stimulation of B16-F10 melanoma cells. Our results show increased levels of the reactive oxygen species after 15 h of melanogenesis stimulation. Following 48 h of melanogenesis stimulation, proliferation was inhibited (by induction of cell cycle arrest in the G1 phase) and the expression levels of p21 mRNA were increased. In addition, melanogenesis stimulation did not induce cellular senescence. Proteomic analysis demonstrated the involvement of proteins from other pathways besides those related to the cell cycle, including protein disulfide isomerase A3, heat-shock protein 70, and fructose biphosphate aldolase A (all up-regulated), and lactate dehydrogenase (down-regulated). In RT-qPCR experiments, the levels of pyruvate kinase M2 mRNA dropped, whereas the levels of ATP synthase (beta-F1) mRNA increased. These data indicate that melanogenesis stimulation of B16-F10 cells leads to alterations in metabolism and cell cycle progression that may contribute to an induction of cell quiescence, which may provide a mechanism of resistance against cellular injury promoted by melanin synthesis.
Departamento de Bioquímica e Biologia Molecular, Setor de Ciências Biológicas, Universidade Federal do Paraná, P.O. Box 19046, CEP 81531-990, Curitiba, PR, Brazil.
Elizabeth S
ES
Cunha
Rebeca
R
Kawahara
Marina K
MK
Kadowaki
Hudson G
HG
Amstalden
Guilhermina R
GR
Noleto
Silvia Maria S C
SM
Cadena
Sheila M B
SM
Winnischofer
Glaucia R
GR
Martinez
D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics; D059585:G1 Phase Cell Cycle Checkpoints; D015870:Gene Expression; D016147:Genes, Tumor Suppressor; D008543:Melanins / Q000096:biosynthesis*; D008546:Melanoma, Experimental / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism; D020543:Proteome / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D014443:Tyrosine / Q000494:pharmacology
C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D008543:Melanins; D009363:Neoplasm Proteins; D020543:Proteome; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D017382:Reactive Oxygen Species; D000643:Ammonium Chloride; D014443:Tyrosine