Macrophages reprogrammed by lung cancer microparticles promote tumor development via release of IL-1β

摘要

L-MPs induced macrophages to upregulate IL-1β expression. a Human PBMC-derived macrophages were treated with HCC827-MPs at a ratio of 1:20 (macrophages: MPs). After 12 h, the cells were collected, and RNA was extracted for real-time PCR analysis of IL-10, arginase 1 (Arg1), VEGF and IL-1β. b, c Human PBMC-derived macrophages were treated with HCC827-MPs at different ratios (cell:MPs, 1:1, 1:5, 1:10, 1:20). RNA, protein and cultured medium were collected after 12, 24, or 72 h of treatment, respectively. Then, the IL-1β expression was analyzed by real-time PCR, western blots (b) or ELISAs (c). dIL-1β mRNA or pro-IL-1β expression of HCC827-MPs, H460-MPs, A549-MPs and Lewis-MPs was analyzed by RT-PCR (left) or western blot (right) analyses. Human PBMC-derived macrophages treated with HCC827-MPs were used as positive controls for A549, HCC827, and H460-MPs. Mouse BMDMs treated with Lewis-MPs were used as a positive control for Lewis-MPs. e Human PBMC-derived macrophages were treated with H460-MPs or A549-MPs for 12 h (left). Mouse BMDMs were treated with Lewis-MPs for 12 h (right). Then, the IL-1β mRNA level was analyzed by real-time PCR. f Human PBMC-derived macrophages were treated with healthy human blood cell-derived MPs at a ratio of 1:20 (cell:MPs). IL-1β mRNA levels were analyzed by real-time PCR (left). BMDMs were treated with wild-type mouse (C57BL/6) blood cell-derived MPs at a ratio of 1:20, and then, the IL-1β mRNA level was analyzed by real-time PCR (right). Error bars indicate the mean ± SEM; n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001