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
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
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DOI：
10.1016/j.yexcr.2012.05.019
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发表时间：
2012-09-10
影响因子：
3.7
通讯作者：
Martinez, Glaucia R.
中科院分区：
文献类型：
--
作者：
Cunha, Elizabeth S.;Kawahara, Rebeca;Martinez, Glaucia R.
考虑到刺激黑素生成可能导致细胞反应的改变，除了黑素生成之外，我们的主要目标是研究黑素生成刺激B16-F10黑色素瘤细胞的细胞效应。我们的研究结果表明，增加水平的活性氧后15小时的黑素生成刺激。黑素生成刺激48小时后，增殖被抑制（通过诱导细胞周期停滞在Cl期），p21 mRNA的表达水平增加。此外，黑素生成刺激不诱导细胞衰老。蛋白质组学分析表明，除了与细胞周期相关的蛋白质外，还涉及其他途径的蛋白质，包括蛋白质二硫键异构酶A3，热休克蛋白70，果糖二磷酸醛缩酶A（均上调）和乳酸脱氢酶（下调）。在RT-qPCR实验中，丙酮酸激酶M2 mRNA水平下降，而ATP合酶（β-F1）mRNA水平升高。这些数据表明，B16-F10细胞的黑素生成刺激导致代谢和细胞周期进程的改变，这可能有助于诱导细胞静止，这可能提供抵抗由黑素合成促进的细胞损伤的机制。(C)2012 Elsevier Inc. All rights reserved.
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 Cl 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. (C) 2012 Elsevier Inc. All rights reserved.