IL‐1β induces MMP‐9 expression via a Ca²⁺‐dependent CaMKII/JNK/c‐JUN cascade in rat brain astrocytes

Abstract

Interleukin (IL)-1beta has been shown to induce matrix metalloproteinase (MMP)-9 expression through mitogen-activated protein kinases, including JNK, in rat brain astrocyte-1 (RBA-1) cells. However, little is known about whether JNK activated by Ca(2+)-dependent CaMKII is associated with MMP-9 expression induced by IL-1beta. Here, we report that the Ca(2+)/CaMKII/JNK/c-Jun participates in the MMP-9 expression induced by IL-1beta. Zymographic, Western blotting, and RT-PCR analyses showed that IL-1beta-induced expression of MMP-9 mRNA and protein was attenuated by Ca(2+) chelator (BAPTA), and the inhibitors of ER Ca(2+)-ATPase (thapsigargin), CaMKII (KN-62), and JNK1/2 (SP600125). IL-1beta also stimulated phosphorylation of CaMKII and JNK1/2, and increase in intracellular Ca(2+) ([Ca(2+)](i)), which were inhibited by pretreatment with BAPTA, thapsigargin (TG), KN-62, or SP600125. Furthermore, the upregulation of MMP-9 protein was blocked by transfection with c-Jun or CaMKII short hairpin RNA (shRNA). We further confirmed that IL-1beta stimulated c-Jun associated with AP-1-binding sites within MMP-9 promoter (-87 to -80 bp and -511 to -497 bp) by immunoprecipitation and chromatin immunoprecipitation (ChIP)-PCR assays. The activation and recruitment of c-Jun to MMP-9 promoter were inhibited by pretreatment with BAPTA, TG, KN-62, or SP600125. Moreover, IL-1beta-induced MMP-9 gene transcription by AP-1 was confirmed by transfection with a MMP-9 promoter-luciferase reporter plasmid with a distal AP-1-binding site (-511 to -497 bp) adjacent to an Ets-binding site-mutation (mt-AP1/Ets-MMP-9). These results demonstrated that in RBA-1 cells, JNK/c-Jun activation was mediated through a Ca(2+)-dependent CaMKII pathway that promoted transcription factor c-Jun/AP-1 recruitment and eventually led to increase in MMP-9 expression by IL-1beta.

Sphingosine 1-phosphate regulates matrix metalloproteinase-9 expression and breast cell invasion through S1P3–Gαq coupling

“…Inflammatory breast cancer is highly metastatic and exhibits an 2226 Journal of Cell Science 124 (13) invasive phenotype both in vivo and in vitro (Charafe-Jauffret et al., 2010;Wu et al, 2009). It has been reported that S1P, a cell membrane-derived phospholipid, plays an important role in the processes of inflammatory responses and cell migration in many cell types (reviewed by Nixon, 2009;Spiegel and Milstien, 2003).…”

“…2+ levels Ca 2+ levels were measured, as described previously (Wu et al, 2009). Cells were washed with PBS, then incubated in DMEM-F-12 containing 5 mM Fura-2/AM for 30 minutes in the dark at 37°C.…”

“…Recent evidence suggests that inflammation is involved in the malignant progression of breast cancer. Invasion, migration and metastasis of breast cancer cells are mediated through NFkB and cytokines such as interleukin 6 (Wu et al, 2009;Barbieri et al, 2010). Inflammatory breast cancer in a human xenograft model displays the phenotype of lymphovascular invasion (Alpaugh et al, 1999).…”

CAMKIIγ suppresses an efferocytosis pathway in macrophages and promotes atherosclerotic plaque necrosis

“…If the resolution process is improved in M-CaMKII-KO lesions, as we predict, this might explain the improvement in protective fibrous cap formation (45). However, it is also possible that M-CaMKII-KO directly inhibits expression or activity of matrix metalloproteinases (see Figure 2E) (46,47), which have been implicated in advanced plaque progression (48,49). Another possible link between inflammation resolution and the findings reported herein is that efferocytosis in general, and MerTK signaling in parNo significant change in Nr1h2 (LXR) was observed.…”

Phosphorylation of moesin by c-Jun N-terminal kinase is important for podosome rosette formation in Src-transformed fibroblasts

“…It is generally believed that JNK promotes cell invasion by activating the transcriptional factor AP1 and the expression of MMPs (Manning and Davis, 2003;Cheung et al, 2006;Wu et al, 2009). Paxillin is known to be required for podosome/ invadopodia formation (Badowski et al, 2008); therefore, we investigated whether JNK plays a role in podosome formation by phosphorylating paxillin.…”