Neutrophilokines and the morphofunctional state of circulating neutrophils in ovarian tumors

Abstract

There is currently no clear understanding of the molecular participants providing cytotoxic and/or cytostatic activity of neutrophils (Nph) in relation to tumor cells. Cytokines produced by neutrophils are necessary for their paracrine and autocrine interactions with surrounding cells. In order to assess the effect of regulatory neutrophilokines on the morphofunctional state of circulating Nph in benign ovarian tumors and ovarian cancer, the ELISA method was used to assess the level of IL-4, IL-6, IL-10, IL-8, IL-18, IFNγ, MCP-1 and MMP-1in neutrophils, expression of CD11b, CD63, CD16, CD95. Determined the rigidity of the membrane and the ability of neutrophils to form NET. Statistical processing of the obtained data was carried out using the software Statistica 13.0, Jamovi 1.6.5.0. It was found that in ovarian cancer the rigidity of neutrophils depends on the level of IL-10, MCP-1, IL-18, IL-8, IL-4, IFNγ, IL-6 in neutrophils. The method of multiple regression revealed the dependence of the ability to form NETs on the level of IL-4 and IL-6 in Nph. Revealed an inverse correlation between the rigidity of the membrane Nph and their ability to form traps in ovarian cancer. In benign ovarian tumors, a noticeable direct correlation was found between the rigidity of the neutrophil membrane and the adhesive marker CD11b. In ovarian cancer, a correlation was found between the rigidity of the Nph membrane and the CD63 degranulation marker. At benign ovarian tumors, no correlations were found between the number of activated neutrophils and the level of intracellular cytokines in Nph. In ovarian cancer, correlations were found between the number of CD11b+Nph and the level of IL-6, IL-8 in them; between the amount of CD63, CD95 and intracellular IL-8. The amount of CD16+Nph correlated with the level of MMP-1 and IL-8 in Nph. The amount of CD95+Nph correlated with the level of IL-18 in Nph. Thus, the change in the level of neutrophilokines in benign ovarian tumors did not correlate with changes in the ability to NETosis, expression of activation markers, but was accompanied by an increase in the rigidity of the membrane of circulating neutrophils. In ovarian cancer, an increase in IL-8 correlated with a decrease in CD16 expression and an increase in CD63; a decrease in CD16 correlated with an increase in MMP-1. An increase in membrane rigidity in ovarian cancer was associated with changes in all considered neutrophilokines (IL-4, IL-6, IL-8, IL-10, IL-18, MCP-1, IFNγ). The combination of IL-4, IL-6, IL-18 indices, NET number and membrane rigidity of circulating Nph (according to the results of multivariate analysis) can be used for differential diagnosis of ovarian cancer.