Phospholipase D Mediates Matrix Metalloproteinase-9 Secretion in Phorbol Ester-stimulated Human Fibrosarcoma Cells

Abstract

Phospholipase D (PLD) has been implicated in vesicle trafficking in the Golgi and hence secretion. In this study, we show that the secretion of matrix metalloproteinase-9 (MMP-9) from HT 1080 human fibrosarcoma cells was stimulated by phorbol 12-myristate 13-acetate in a time-and dose-dependent manner that involved protein kinase C. The phorbol ester also increased PLD activity in the cells. Evidence that PLD was involved in the stimulation of MMP-9 secretion was provided by the observations that the secretion of MMP-9 was stimulated by the introduction of short-chain phosphatidic acid (PA) into the growth medium and that inhibition of PA production by 1-propanol inhibited secretion. Using a short-chain diacylglycerol we excluded the possibility that MMP-9 secretion was induced by diacylglycerol formed from PA by phosphatidic acid phosphatase. Furthermore, propranolol, an inhibitor of this enzyme, had no effect on secretion induced by either phorbol 12-myristate 13-acetate or PA. The data presented here indicate that activation of protein kinase C increases MMP-9 secretion in HT 1080 cells and implicate PLD and PA formation in the effect.

RalA requirement for v-Src- and v-Ras-induced tumorigenicity and overproduction of urokinase-type plasminogen activator: involvement of metalloproteases

“…These data suggest that the increased expression of MMP in the v-Srctransformed cells is dependent upon RalA and the elevated speci®c activity observed in the v-Rastransformed cells is independent of RalA. Exton and co-workers recently reported that MMP-9 secretion induced by phorbol esters was dependent on PLD and phosphatidic acid production (Williger et al, 1999). It is unlikely that the mechanism was the same as that reported here since phorbol esters activate PLD by activating protein kinase C, which can activate PLD directly (Exton, 1998).…”

Choline in acute coronary syndrome: an emerging biomarker with implications for the integrated assessment of plaque vulnerability

“…Corresponding with this concept, very high levels of WBCho in combination with normal PCho have been found in cases with subacute stent thrombosis, sudden death and massive intravascular thrombi [26]. Another potential mechanism responsible for the activation of PLD in the monocyte-macrophage system is the involvement of PLD in the secretion of matrix metallo proteinases [27], which are crucial for causing plaque rupture. Furthermore, a change of PLD expression has been demonstrated in human plaque macrophages being colocalized with oxidation epitopes [28].…”

Multiple signaling pathways involved in activation of matrix metalloproteinase-9 (MMP-9) by heregulin-β1 in human breast cancer cells

“…Involvement of JNK signaling pathway in regulation of MMP-9 is also reported (Gum et al, 1997). Finally, secretion of newly synthesized MMP-9 from HT1080 human ®brosarcoma cells was regulated by phospholipase D activity (Williger et al, 1999), which may regulate MMP-9 production at a late stage.…”