TonEBP expression is essential in the IL-1β–induced migration and invasion of human A549 lung cancer cells

Figures

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  Figure 1. IL-1β secretion and expression in A549 cocultured-MΦ (A) IL-1β levels were determined via ELISA. (B) IL-1β and GAPDH expression were determined from whole-cell lysates. (C) IL-1β mRNA expression was analyzed via quantitative RT-PCR. Data are presented as the mean ± SEM of three independent experiments (***p < 0.001, compared with monocytes).

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  Figure 2. IL-1β secretion by A549 cocultured-MΦ promotes cancer cell migration and invasion. (A–D) A549 cells were treated with IL-1β. (E–H) A549 cells were incubated with A549 cocultured-MΦ-conditioned media in the presence or absence of IL-1RA for 24 h. Wound healing and transwell migration assays were conducted to determine the effects of IL-1β on cancer cell migration and invasion. Data are presented as the mean ± SEM of three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001, compared with 0 ng/mL and control; NI: non-induced).

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  Figure 3. Effects of MAPK and NF-κB inhibitors on IL-1β–induced cancer cell migration and invasion. (A) p-p38/t-p38, p-ERK/t-ERK, and IκBα expression levels were determined using whole-cell lysates. (B, C) A549 cells were pretreated with ERK, p38 and IKK2 inhibitors for 1 h and then treated with IL-1β for 24 h. The effects of MAPK and NF-κB inhibitors on cancer cell migration and invasion were determined using a transwell migration assay. Data are presented as the mean ± SEM of three independent experiments (***p < 0.001, compared with the -).

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  Figure 4. Inhibitory effects of IPA3 on IL-1β–induced migration and invasion of A549 cells. p-PAK1/t-PAK1 expression levels were determined using whole-cell lysates. (A, B) Western blot results. (C–E) A549 cells were incubated with IL-1β in the presence or absence of IPA3 for 24 h. The effects of IPA3 on cancer cell migration and invasion were determined via wound healing and transwell migration assays. (F) A549 cells were pretreated with ERK, p38 and IKK2 inhibitors for 1 h and then treated with IL-1β for 24 h. Data are presented as the mean ± SEM of three independent experiments (*p < 0.05, compared with the con and -; con: control).

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  Figure 5. Effects of TonEBP depletion on cancer cell migration and invasion. A549 cells were transfected with siTonEBP or siNC for 24 h and then treated with IL-1β for 24 h. (A–D) Migration and Invasion were quantified via wound healing and transwell migration assays. Data are presented as the mean ± SEM of three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001, compared with the siNC treatment; NC: negative control).

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  Figure 6. Expression of p-PAK1 and T-PAK1 on TonEBP deficiency. (A) TonEBP, p-p38/t-p38, p-ERK/t-ERK, and IκBα expression levels were determined using whole-cell lysates. A549 cells were transfected with siTonEBP or siNC for 48 h and then treated with IL-1β for 1 h. (B) PAK1 mRNA expression was analyzed via quantitative RT-PCR. (C) p-PAK1 and T-PAK1 expression was determined using whole-cell lysates. Data are presented as the mean ± SEM of three independent experiments (**p < 0.01, ***p < 0.001, compared with the siNC treatment; NC: negative control).

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  Figure 7. Schematic diagram of the effects of IL-1β and TonEBP expression on cell migration and invasion in A549 cells.