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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: cPLA2 blockade attenuates S100A7-mediated breast tumorigenicity by inhibiting the immunosuppressive tumor microenvironment

Fig. 4

S100A7/cPLA2 signaling enhances the infiltration of tumor-associated macrophages (TAMs) through increased secretion of PGE2 (A). Schematic representation showing the treatment of conditioned media (CM) derived from MDA-MB-231 vector (231 V) and S100A7 overexpressing cells (S7OE-231) in presence or absence of PGE2 receptor, EP4 inhibitor on migration of bone marrow derived macrophages (BMDMs). Bar diagram shows the number of migrated BMDMs per field stimulated with CM derived from (B). 231 V and S7OE-231, (C). MVT1 cells treated with VC or AF3 or BMDMs preincubated with EP4 receptor antagonist (5 μM L161,982) for 2 h before stimulation with CM of MVT1 cells. Overnight migration using transwell inserts was performed. The bars indicate the means ± SEMs of 3 replicates (D). Flow cytometric analysis of MHC-II and CD206 in BMDMs cultured in CM of 231 V and S7OE-231 cells or BMDM preincubated with EP4 receptor antagonist (5 μM L161,982) for 2 h before stimulation with CM of S7OE-231 cells (E). Quantitative analysis of blood PGE2 (pg/mL) in MVT1 tumor-bearing Tet-off (normal diet) and Tet-on (DOX diet) mice treated with VC or AACOCF3 (AF3) (F). CD11b+F4/80+ TAMs (out of CD45+ cells) in tumors harvested from MVT1 tumor-bearing Tet-off (normal diet) and Tet-on (DOX diet) mice treated with VC or AF3 (G). The bar diagram indicates the means ±SEMs of three replicates (H). Representative IHC images of CD163+ M2-TAMs in adjacent normal, low-grade, and high-grade invasive breast cancer specimens (source: Biomax) (I). Box plot showing the percentage (%) of CD163+ M2-TAMs in normal, low-grade, and high-grade invasive breast cancer specimens. Non-parametric test (Independent-Samples Median Test) for comparing high grade to low grade and adjacent normal specimens of breast cancer patients. Bonferroni correction was applied for multiple tests. Spearman rho’s correlation coefficient analysis between abundance of CD163+ M2-TAMs with protein levels (% high positive) of (J). S100A7 and (K). cPLA2 in TMAs of breast cancer patients (n = 70) (source: Biomax) (L). Spearman rho’s correlation coefficient analysis of cPLA2 mRNA (PLA2G4A) expression with infiltration of M2-TAMs in breast cancer patients (source: TIMER2.0; http://timer.cistrome.org/). ns: non-significant, *P < 0.05, ** P < 0.01, *** P < 0.001. One way ANOVA was used for multiple group comparisons

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