Mouse Melanoma Cell Migration is Dependent on Production of Reactive Oxygen Species under Normoxia Condition

Abstract

Cell migration plays a role in many physiological and pathological processes. Reactive oxygen species (ROS) produced in mammalian cells influence intracellular signaling processes which in turn regulate various biological activities. Here, we investigated whether melanoma cell migration could be controlled by ROS production under normoxia condition. Cell migration was measured by wound healing assay after scratching confluent monolayer of B16F10 mouse melanoma cells. Cell migration was enhanced over 12 h after scratching cells. In addition, we found that ROS production was increased by scratching cells. ERK phosphorylation was also increased by scratching cells but it was decreased by the treatment with ROS scavengers, N-acetylcysteine (NAC). Tumor cell migration was inhibited by the treatment with PD98059, ERK inhibitor, NAC or DPI, well-known ROS scavengers. Tumor cell growth as judged by succinate dehydrogenase activity was inhibited by NAC treatment. When mice were intraperitoneally administered with NAC, the intracellular ROS production was reduced in peripheral blood mononuclear cells. In addition, B16F10 tumor growth was significantly inhibited by in vivo treatment with NAC. Collectively, these findings suggest that tumor cell migration and growth could be controlled by ROS production and its downstream signaling pathways, in vitro and in vivo.

Characterization of preneoplastic and neoplastic rat mesothelial cell lines: the involvement of TETs, DNMTs, and 5-hydroxymethylcytosine

“…Cell migration was analyzed for all rat cell lines by the scratching test [71]. When cell density was confluent, for each cell line cultivated in 12-well plates (3.5 cm 2 / well), one wound line in the form of a cross was made by scratching the cell monolayer with a plastic pipette tip.…”

The role of chemical elements in melanoma

“…The antioxidant pattern has been found altered in experimental melanoma (Lazescu, 2013) and in simvastatin-treated melanoma cells undergoing a p53/p21 -mediated senescence (Guterres, 2013); the ROS pool can be produced by NADPH oxidase enzymes (NOX family), representing a promising target for melanoma treatment (LiuSmith, 2014). ROS have been found to mediate the pro-angiogenic potential of melanoma cells (Schaafhausen, 2013;Vartanian, 2007); ROS mediate the in vitro apoptosis and growth inhibition of A375 human melanoma cells and other melanoma cells, induced by several different agents (Huang, 2014;Huang, 2012;Chakraborty, 2013;Ghosh, 2013;Mayola, 2011;Zhang, 2010;Morrison, 2010;Chou, 2009;Wang, 2008;Verhaegen, 2006) and of B16F10 mouse melanoma cells migration properties (Im, 2012). Blocking ROS production has been shown to reduce the anti-5 melanoma activity of Cucurbitacin B (Zhang 2011) while decreasing ROS promotes melanoma growth (Huang 2010).…”

Quantifying rates of cell migration and cell proliferation in co-culture barrier assays reveals how skin and melanoma cells interact during melanoma spreading and invasion

“…Therefore, to understand how quickly a population of melanoma cells spreads through the surrounding environment, it is important to develop techniques that allow us to quantify the rates of cell migration and cell proliferation (Treloar et al, 2013). Previous in vitro studies examining the spatial spreading of populations of melanoma cells have focused on monoculture experiments that contain only melanoma cells (Cornil et al, 1991; Im et al, 2012; Justus et al, 2014; Treloar et al, 2013). To make these kinds of in vitro studies more relevant to the in vivo situation, it is important to investigate, and quantify, how the spatial spreading of melanoma cells is affected by interactions with other cells types, such as fibroblasts.…”