S100A9 Regulated M1/M2 Macrophage Polarization in Interleukin-10-Induced Promotion of Malignant Pleural Effusion

Figure 4

S100A9 knockdown in mice suppressed the development of MPE and regulated M1/M2 macrophage polarization in MPE. (a) S100A9 was knocked down by the application of Chol-siS100A9 in WT mice, and the downregulation of S100A9 in MPEs, blood, and spleens was verified by qRT–PCR (each n = 5). (b) Comparisons of MPE volume and intrapleural tumors in mice receiving Chol-siS100A9 and cholesterol-conjugated siCtr (Chol-siCtr). (c) Representative flow cytometric dot plots (left panel) and statistical comparisons (right panel) of MPE macrophages of the S100A9-knockdown and control groups (each n = 6). (d) Representative flow cytometric dot plots (left panel) and statistical comparisons (right panel) of M2 and M1 macrophages in MPEs from the S100A9-knockdown and control groups (each n = 6). (e) Transwell migration assay was performed on MC38 cells treated with macrophages from MPEs of the S100A9-knockdown and control groups; the representative images (upper panel) and statistically significant numbers of migrated cells (lower panel) are shown (n = 4). (f) Representative flow cytometric dot plots (upper panel) and comparisons (lower panel) of apoptosis in MC38 cells cocultured with macrophages from MPEs of the S100A9-knockdown and control groups (n = 4). (g) Correlation analysis of IL-10 and S100A9 gene expression was performed by GEPIA (data were from Kaplan–Meier plotter, http://gepia.cancer-pku.cn/index.html). (h) Overall survival of ungrouped lung cancer patients (left panel) and lung adenocarcinoma patients (right panel) with high- or low-S100A9 expression was evaluated using the Kaplan–Meier survival curves and compared using the pairwise log-rank tests (data were from Kaplan–Meier plotter, http://kmplot.com/analysis/). Data are presented as mean ± SD. , , , compared by Student’s t test.

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