Oxidative Stress Differentially Influences the Survival and Metabolism of Cells in the Melanoma Microenvironment

Abstract

The cellular composition of the tumor microenvironment, including tumor, immune, stromal, and endothelial cells, significantly influences responses to cancer therapies. In this study, we analyzed the impact of oxidative stress, induced by cold atmospheric plasma (CAP), on tumor cells, T cells, and macrophages, which comprise part of the melanoma microenvironment. To accomplish this, cells were grown in different in vitro cell culture models and were treated with varying amounts of CAP. Subsequent alterations in viability, proliferation, and phenotype were analyzed via flow cytometry and metabolic alterations by Seahorse Cell Mito Stress Tests. It was found that cells generally exhibited reduced viability and proliferation, stemming from CAP induced G2/M cell cycle arrest and subsequent apoptosis, as well as increased mitochondrial stress following CAP treatment. Overall, sensitivity to CAP treatment was found to be cell type dependent with T cells being the most affected. Interestingly, CAP influenced the polarization of M0 macrophages to a “M0/M2-like” phenotype, and M1 macrophages were found to display a heightened sensitivity to CAP induced mitochondrial stress. CAP also inhibited the growth and killed melanoma cells in 2D and 3D in vitro cell culture models in a dose-dependent manner. Improving our understanding of oxidative stress, mechanisms to manipulate it, and its implications for the tumor microenvironment may help in the discovery of new therapeutic targets.

GARP Regulates the Immune Capacity of a Human Autologous Platelet Concentrate

“…Buffy coats were obtained from healthy volunteers, and macrophages were isolated from them as previously described [ 31 ]. In brief, 150 million peripheral blood mononuclear cells (PBMC) were plated in RPMI1640 (Thermo Fisher Scientific, #31870, Waltham, MA, USA), containing 1% Glutamax (Thermo Fisher Scientific, #35050038) and 0.1% Primocin (InvivoGen, #ant-pm-2, San Diego, CA, USA) in cell culture Petri dishes (Corning, #430167, Corning, NY, USA) for at least 1 h in a 37 °C 5% CO 2 incubator to promote attachment.…”

“…Adherent cells were cultured in the medium described above containing an additional 1% heat inactivated human plasma and 50 ng/mL M-CSF (ImmunoTools, #11343113, Friesoythe, Germany). Generation of heat inactivated human plasma was prepared as previously described [ 31 ]. After 5–7 days, “M0” macrophages were collected with Accutase (Thermo Fisher Scientific, #00-4555-56) and replated at 10 6 cells/well in 6 well plates (Corning, #3506).…”

“…After 5–7 days, “M0” macrophages were collected with Accutase (Thermo Fisher Scientific, #00-4555-56) and replated at 10 6 cells/well in 6 well plates (Corning, #3506). Cells underwent polarization to an “M1-like” or a “M2-like” phenotype as previously described [ 31 ]. For polarization experiments, “M0-like” macrophages were treated with 400 µL of iPRF/well.…”

Plasma‐activated medium triggers immunomodulation and autophagic activity for periodontal regeneration

“…Activated plasma‐polarized macrophages can inhibit cell viability, EMT, and cancer stemness in glioblastoma multiforme 43 . In 2022, Trzeciak discovered that M0 resting macrophages treated with CAP exhibited an increase in the M2 phenotype, focusing on the effects and changes of plasma‐induced oxidative stress on macrophages in the melanoma tumor microenvironment 44 . Different plasma treatment parameters and application scenarios, as shown in Table 2, may produce opposite outcomes.…”

Phototoxicity and cell passage affect intracellular reactive oxygen species levels and sensitivity towards non-thermal plasma treatment in fluorescently-labeled cancer cells

“…Live-cell fluorescence imaging has been a valuable tool in the field of plasma medicine for investigating cellular and subcellular NTP treatment effects for different biomedical applications, including wound healing and cancer therapy [13][14][15][16]. Mounting evidence has demonstrated that the versatile medicinal properties of NTP arise from the diverse short-lived and persistent reactive species (e.g.…”