Fig. 6
IL30-dependent regulation of CXCL5 production in human PC cells and survival curves of PC patients based on the expression of IL30 and SOCS3 in their clinical samples. A, B Elisa assay of CXCL5 release by wild type DU145 (A) and PC3 (B) cells, after the treatment with anti-IL30 Abs. A ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with untreated DU145 cells. **p < 0.01, Tukey HSD test compared with DU145 cells untreated or treated with 5 μg/mL. B ANOVA: p < 0.001. *p < 0.01, Tukey HSD test compared with untreated PC3 cells. Results are expressed as mean ± SD. C, D Elisa assay of CXCL5 release by wild type, EV- and IL30 gene-transfected DU145 (C) and PC3 (D) cells. ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with WT and EV-transfected DU145 cells. Results are expressed as mean ± SD. E Cytofluorimetric analyses of CXCR2 expression in PC3 and DU145 cells. Red lines: isotype control. Experiments were performed in triplicate. F, G MTT assay of DU145 (F) and PC3 (G) cells, untreated (0.0 μg/mL) or treated with anti-CXCL5 Abs (0.2, 0.5, 1.0 μg/mL in DU145; 0.25, 0.80, 1.50 μg/mL in PC3). (F) ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with 0.0 and 0.2 μg/mL. (G) ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with 0.0 µg/mL. **p < 0.01, Tukey HSD test compared with 0.0 and 0.25 μg/mL. Results are expressed as mean ± SD. H, I MTT assay of DU145 (H) and PC3 (I) cells, untreated (0.0 ng/mL) or treated with rhCXCL5 (5, 7, 10, ng/mL in DU145; 5, 10, 30, 50 ng/mL in PC3). H ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with 0 ng/mL. **p < 0.05, Tukey HSD test compared with 0 and 5 ng/mL. I ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with 0 ng/mL. **p < 0.01, Tukey HSD test compared with 0 and 5 ng/mL. ***p < 0.01, Tukey HSD test compared with 0, 5 and 10 ng/mL. Results are expressed as mean ± SD. J, K MTT assay of wild type and IL30 gene-transfected DU145 (J) and PC3 (K) cells, untreated (0.0 μg/mL), or treated with anti-CXCL5 Abs (0.5 μg/mL and 0.80 μg/mL, respectively). ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with wild type cells. **p < 0.01, Tukey HSD test compared with wild type and IL30 gene-transfected cells. Results are expressed as mean ± SD. L Kaplan–Meier curves representing, for each time point, the fraction of surviving PC patients, from the PanCancer collection, classified, based on mRNA expression levels in tumor samples, as IL30 mRNAHigh (n.16/494) and IL30 mRNAMod (n.478/494). Log-rank test, p = 0.01. M Kaplan–Meier curves representing, for each time point, the fraction of surviving PC patients, from the PanCancer collection, classified, based on mRNA expression levels in tumor samples, as SOCS3 mRNAMod (n.476/493) and SOCS3 mRNALow (n.17/493). Log-rank test, p = 0.04. N Expression of IL30 (a, b) and SOCS3 (c, d) in PC tissues obtained from patients with (IL30PosSOCS3Neg) or without (IL30NegSOCS3Pos) biochemical recurrence (BCR). Magnification: × 630. O Kaplan–Meier curves representing, for each time point, the fraction of surviving PC patients classified, based on IL30 and SOCS3 expression in both tumor cells and infiltrating leukocytes, as IL30PosSOCS3NegPC (n.29), IL30PosSOCS3PosPC (n.18), IL30NegSOCS3NegPC (n.49) and IL30NegSOCS3PosPC (n. 67). Log-rank test, p < 0.001