The Macrophage Activator GcMAF-RF Enhances the Antitumor Effect of Karanahan Technology through Induction of M2–M1 Macrophage Reprogramming

Figure 3

Analysis of synthesis of specific pro- and anti-inflammatory factor mRNAs in reducing tumors isolated from tumor-bearing mice and mice treated with Karanahan technology and in synergy with GcMAF-RF. (a) Relative levels of mRNA expression of some pro- and anti-inflammatory factor genes in populations of tumor cells isolated from mice treated with Karanahan technology and in synergy with GcMAF-RF compared with cells isolated from tumor-bearing mice (their expression level = 1; the Y-axis is logarithmic). Box and whisker plot; the black asterisk denotes the significant difference with control (tumor-bearing mice); the hash indicates the significant difference between experimental Karanahan and Karanahan + GcMAF-RF groups, ; Mann–Whitney U test. (b) Graphical representation of the relative levels of mRNA expression of some pro- and anti-inflammatory factor genes in peritoneal macrophages (PMs) and cells of tumor-associated stroma (TAS) isolated from tumor-bearing mice, mice treated with Karanahan technology, and mice treated with Karanahan technology in synergy with GcMAF-RF. (b1) The figure schematically shows peritoneal macrophages and TAS macrophages of tumor-bearing mice. Blue color denotes the M2 anti-inflammatory phenotype; white color denotes the normal M0 phenotype. A factor (cytokine) name without an arrow means its presence in a sample. A crossed-out cytokine name means its absence in a sample. The presence/absence in a sample was assessed by polymerase chain reaction with specific primers. (b2) Evaluation of peritoneal macrophages and TAS cells phenotype changes after treatments. The figure schematically shows peritoneal macrophages and TAS macrophages of mice treated with Karanahan technology alone and in synergy with GcMAF-RF. An increase/decrease in the number of schematic cells after treatments indicates an increase/decrease in the number of CD11b+Ly-6C+ monocytic cells among peritoneal macrophages and TAS cells. Blue color denotes the M2 anti-inflammatory phenotype; red color denotes the M1 proinflammatory phenotype. Relative levels of mRNA expression of pro- and anti-inflammatory factor (cytokine) genes in peritoneal macrophages and TAS cells of mice treated with Karanahan technology alone and in synergy with GcMAF-RF compared with those in peritoneal macrophages and TAS cells of tumor-bearing mice were assessed by Real-time PCR. A factor (cytokine) name with an up (down) arrow means an increase (decrease) in a relative level of factor (cytokine) mRNA expression in cells isolated from treated mice compared with that in cells isolated from tumor-bearing mice. A factor (cytokine) name with bidirectional arrows means multidirectional changes in a relative level of the factor mRNA in cells isolated from treated mice compared with that in cells isolated from tumor-bearing mice. The bold arrow indicates additive inhibition of TGF-β1 expression under the synergistic action of Karanahan technology and GcMAF-RF.

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