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Fig. 4 | Molecular Cancer

Fig. 4

From: PKN2 in colon cancer cells inhibits M2 phenotype polarization of tumor-associated macrophages via regulating DUSP6-Erk1/2 pathway

Fig. 4

PKN2 negatively regulates IL4 and IL10 productionin colon cancer cells. a Gene expression profiles analysis was done in PKN2-K686R, PKN2-WT stably overexpressed or wild-type HCT116 cells. Genes in KEGG ‘chemokine signaling pathway’ and ‘cytokine-cytokine receptor interaction’ clusters showing 2-fold or higher differential expression were selected. b The clustered heatmap of two cytokine genes IL4 and IL10 were identified from PKN2-WT and PKN2-K686R HCT116 cells. The color-coding applies to gene expression level (log2) with 0 as a median. c The mRNA level of IL4 and IL10 in WT, PKN2-K686R and PKN2-WT HCT116 cells was assessed using RT-PCR.*, P < 0.05; ***, P < 0.001 versus WT. d Stable clone of SW480, HCT116 and HT-29 cells (as indicated in Fig. 3) were cultured for 48 h. The level of IL10 and IL4 in the culture supernatants was assessed using ELISA.*, P < 0.05; **, P < 0.01 versus Vector or shCTL. e SW480 cells were stably infected with shPKN2-1, shPKN2-2, PKN2-WT or vector lentivirus. Luciferase assays were performed to detect transcription factor binding activity of IL4 and IL10. Relative fold-change in luciferase activity was shown. *, P < 0.05; **, P < 0.01 versus WT. f Human CD14+ monocytes were cocultured with stably shCTL or shPKN2 transfected HT-29 with/without neutralizing antibody of IL10 (2 μg/ml) or IL4 (0.5 μg/ml) for 4 days. Surface expression of CD206 in differentiated macrophages was detected using flow cytometry. ***, P < 0.001 versus shCTL. #, P < 0.05; ##, P < 0.01 versus shPKN2. g Human CD14+ monocytes were cocultured with HCT116 stably preinfected with vector or PKN2-WT with/without neutralizing antibody of IL10 (2 μg/ml) or IL4 (0.5 μg/ml) for 4 days. Surface expression of CD86 was detected using flow cytometry.***, P < 0.001 versus Vector. #, P < 0.05 versus PKN2-WT

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