Tumor-suppressor Lkb1 inactivation promotes neutrophil accumulation via proinflammatory cytokines and chemokines. Tumor-suppressor Lkb1 inactivation promotes.

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Tumor-suppressor Lkb1 inactivation promotes neutrophil accumulation via proinflammatory cytokines and chemokines. Tumor-suppressor Lkb1 inactivation promotes neutrophil accumulation via proinflammatory cytokines and chemokines. A, immune cell populations in the lung tumors from Kras (K) and Kras/Lkb1 (KL) mouse models. Representative flow cytometry data (live/single/total CD45+ cells) from each mouse model (left). Total counts of TAN: CD11b+Ly-6G+ cells and TAMs: CD11c+CD11b−CD103− from Kras (K; n = 8) and Kras/Lkb1 (KL; n = 8) mice. **, P < 0.01; ***, P < 0.001. B, neutrophil counts in the spleen and peripheral blood from Kras (n = 8) or Kras/Lkb1 (n = 8) mice (right). **, P < 0.01. C, expression of immune-modulating factors from RNA sequencing of the sorted tumor cells (CD45−EpCAM+) in Kras (K Ep) or Kras/Lkb1 (KL Ep) mice and uninduced normal lung CD45−EpCAM+ cells (control). Each column consists of a combination of samples derived from 3 to 4 mice. Log-transformed FPKM values are shown, colored blue/red for low/high expression, respectively. Epcam and Cd45 expression are shown as positive and negative controls. Differential expression is shown as fold-change values, colored blue/red for under/overexpression compared with controls. D and E, chemokine and cytokine levels in the culture supernatants after 48-hour incubation from Kras/p53 (KP; n = 3) versus Kras/p53/Lkb1 (KPL; n = 3) cell lines generated from mouse lung tumors; ***, P < 0.001. Data indicate three replicate wells and are representative of three independent experiments (D) and BALF from littermate controls (n = 5), Kras mice (n = 8) or Kras/Lkb1 mice (n = 8; E). *, P < 0.05; **, P < 0.01 . F, Western blot analysis for pSTAT3, STAT3 levels in Kras versus Kras/Lkb1 tumors. Each column represents tumor from a different mouse and actin represents loading control. G, IL1α level in the BALF from C (n = 5), Kras (n = 8), or Kras/Lkb1 (n = 8) mice. *, P < 0.05. H, IL6 levels in culture supernatants measured 24 hours after IL1α stimulation (0, 5, and 20 ng/mL) of KP (n = 3) versus KPL (n = 3) cell lines. Data indicate three replicate wells and are representative of three independent experiments. Shohei Koyama et al. Cancer Res 2016;76:999-1008 ©2016 by American Association for Cancer Research