Exogenous C2 Ceramide Suppresses Matrix Metalloproteinase Gene Expression by Inhibiting ROS Production and MAPK Signaling Pathways in PMA-Stimulated Human Astroglioma Cells

Abstract

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases, which play a pivotal role in invasion, migration, and angiogenesis of glioma. Therefore, controlling MMPs is potentially an important therapeutic strategy for glioma. In the present study, we found that exogenous cell-permeable short-chain C2 ceramide inhibits phorbol myristate acetate (PMA)-induced MMP-1, -3, and -9 gene expressions in U87MG and U373MG human astroglioma cells. In addition, C2 ceramide inhibited the protein secretion and enzymatic activities of MMP-1, -3, and -9. The Matrigel invasion assay and wound healing assay showed that C2 ceramide suppresses the in vitro invasion and migration of glioma cells, which appears to be involved in strong inhibition of MMPs by C2 ceramide. Subsequent mechanistic studies revealed that C2 ceramide inhibits PMA-induced mitogen-activated protein kinase (MAPK) phosphorylation and nuclear factor (NF)-κB/activator protein (AP)-1 DNA binding activities. Furthermore, C2 ceramide significantly inhibited PMA-induced reactive oxygen species (ROS) production and NADPH oxidase 4 (NOX4) expression, and inhibition of ROS by diphenylene iodonium (DPI, NADPH oxidase inhibitor) mimicked the effects of C2 ceramide on MMP expression and NF-κB/AP-1 via inhibition of p38 MAPK. The results suggest C2 ceramide inhibits MMP expression and glioma invasion, at least partly, by modulating ROS-p38 MAPK signaling axis and other MAPK signaling pathways.

Acid Sphingomyelinase Down‐regulation Alleviates Vascular Endothelial Insulin Resistance in Diabetic Rats

“…It has been shown that ASM and ceramide could activate the Nox2 isoform in non‐muscle cells through enhancing phosphorylation of p47 phox 44 . Moreover, Nox4‐dependent O 2 −· production induced by PA was associated with ceramide in human astroglioma cells . We demonstrated that ASM down‐regulation inhibited the protein expression of Nox2 and Nox4 via ceramide to alleviate insulin resistance in PA‐induced RAECs.…”

Rhein suppresses matrix metalloproteinase production by regulating the Rac1/ROS/MAPK/AP-1 pathway in human ovarian carcinoma cells

“…. ROS contribution to cell migration and invasion has been well documented (22). We therefore, speculated that rhein might suppress SKOV3-PM4 cell migration via scavenging ROS.…”

“…The migration of cancer cells is pivotal for cancer invasion and metastasis (10,11,22). Cancer malignancy is proportional to the ability of cancer cell migration.…”

“…It was reported that transcription factor AP-1 is regulated by MAPK and AP-1 is also known to be a key transcription factor of MMPs (22,25,26). Woo et al reported that activation of one or more MAPK pathways was often associated with the MMP expression in various cell types (26).…”

NOX4, a new genetic target for anti‐cancer therapy in digestive system cancer

“…In glioblastomas, NOX4‐generated ROS are essential for hypoxia‐induced progression through the activation of NF‐κB and the ERK‐mediated stimulation of matrix metalloproteinase‐9 . The use of the NOX4 inhibitor diphenylene iodonium (DPI) to interrupt the ROS/MAPK signaling axis decreased matrix metalloproteinase gene expression, NF‐κB and AP‐1 reporter gene activities, and glioma invasion . In the metabolism of tumor cells, NOX4‐induced glycolysis supports cloned tumor cell growth via MYC upregulation, which is dependent on ROS/PI3K/AKT signaling and subsequently contributes to the malignant progression of non‐small cell lung carcinoma .…”