Cold atmospheric plasma-modulated phorbol 12-myristate 13-acetate-induced differentiation of U937 cells to macrophage-like cells

Abstract

Monocytes are recruited to injured tissue sites and differentiate into tissue macrophages or dendritic cells to protect against pathogens and repair the damaged tissues. Phorbol-12-myristate-13-acetate (PMA) is a well-known stimulus commonly used for differentiation of monocytes into macrophage-like cells (MϕLC). Here, we report the effect of Cold Atmospheric Plasma (CAP) on PMA-induced U937 differentiation into MϕLC. Treatment of U937 cells with PMA for 3 days and resting for 4 days increased the size of cytoplasm as compared with nucleus, and exposure to CAP before addition of PMA led to further increase in cytoplasm indicating the ability of CAP to modulate the differentiation of monocytes. Exposure of U937 cells to CAP or PMA increased cellular reactive oxygen species (ROS) level and the combination led to further augmentation of ROS. Treatment of U937 cells with PMA displayed a biphasic activation of proinflammatory transcription factor NF-κB, which plays an important role in differentiation and pretreatment with CAP further increased PMA induced NF-κB-DNA-binding activity. CAP also increased lipopolysaccharide induced secretion of TNF-α and IL-6 in MϕLC. Further investigation revealed that MϕLC or CAP-treated MϕLC were more resistant to anticancer drugs such as doxorubicin and 5-fluorouracil (5-FU) than U937 cells. Our present studies suggest an alternate protocol to modulate the differentiation of U937 cells into MϕLC by combining CAP and PMA.